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Magnetic Beads DNA Purification: History and Recent Developments

Nucleic acid (DNA and RNA) purification and amplification is an important tool for molecular biology and an important step before many biochemical and diagnostic processes. These techniques have made great progress recently[1] [2] due to the increasing number of sudden and public health-threatening infectious diseases (e.g. Ebola virus, Zika virus and more recently SARS-Covid). For quickly and reliably diagnosing these diseases, nucleic acid detection are key tools used in on-site immunological technologies and rapid test kits (for magnetic mRNA purification refer to “Oligo dT-coated magnetic beads: the benefits of their application for mRNA purification”).

 

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Protein Isolation protocol

A protein isolation protocol aims to safely and efficiently separate a specific protein from a complex biological sample. Isolation of proteins can be performed on mammalian, insect, plant, yeast, or bacteria samples. The first step of a protein isolation protocol requires lysing cells within a sample to reveal their contents which can include DNA, RNA, organelles, and various proteins. There are several methods of cell lysing which range from chemical to physical perturbation of a cell membrane or wall. You can read more about cell lysis in this article:How to lyse cells. After cells are lysed, protein purification can be performed. Two popular methods include affinity chromatography and magnetic bead separation

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Sandwich Hybridization: DNA/RNA Capture using Magnetic Beads

Sandwich DNA/RNA hybridization is a technique designed for the detection and quantitation of nucleic acids within crude biological samples. DNA/RNA sandwich hybridization has also widely been used not only to identify a specific DNA/RNA  sequence, but also to distinguish single nucleotide polymorphisms (SNPs) between wild-type and mutant DNA/RNA.

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Magnetic Immunoprecipitation and co-Immunoprecipation and IP Input Analysis

Immunoprecipitation (IP) is a technique for capturing specific proteins from a complex solution via antibody-antigen affinity. In IP the goal is to target and isolate a specific protein, whereas in co-IP the goal is to identify protein complexes and other macromolecules bound to the target in the sample solution. The targeted protein complexes can later undergo analysis to identify specific binding partners, determine binding affinities, or study the kinetic relationship between binding and the function of the target protein. Recently, magnetic nanoparticle technologies have greatly advanced co-IP experiments. Magnetic co-IP protocols offer simple, ultra-fast workflows, versatility, and help generate highly concentrated complexes. After performing a magnetic co-IP protocol, the protein capture efficiency can be measured by IP input, which is essentially the total protein lysate and what is eluted from the magnetic beads after. The IP is often tested through SDS-PAGE and Western blot analysis.

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E-book launch - Standardization of Magnetic Beads Separation Processes

A Guide for Establishing Robust SOP

As the applications of magnetic beads continue to expand, many scientists find themselves navigating magnetic separation protocols for the first time. While successfully employing magnetic beads at the R&D scale, the transition to larger-scale manufacturing can be intricate.

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Synthesis of Magnetic Nanoparticles: Protection, Functionalization, and Application

Magnetic nanoparticles have risen in popularity in medical and biotechnology fields over the past decade. The ideal magnetic properties of nanoparticles for biomedical applications are that they exhibit a form of magnetism called superparamagnetism, where they can be magnetized by an externally applied magnetic field and quickly returned to a non-magnetic state once the field is removed. This feature also prevents magnetic nanoparticles from exhibiting an active behavior when there is no applied field.

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Protein Purification Buffer

The Importance of protein purification buffer

A buffer, by definition, helps a solution to resist changes in pH when small quantities of acid or base are added to it.Protein purification protocols utilize various buffers to aid in stabilization at each step of the process. For example, lysis buffers are initially used to establish an ionic strength in the solution, and binding buffers help in target proteins bind to an immobilizing support matrix, like magnetic beads.

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Magnetic Protein Expression and Purification

In essence, protein expression and purification methods aim to produce functional proteins to deduce the structural, physical, and chemical attributes of a protein for  biomedical research. Protein expression involves understanding the way in which proteins are synthesized, modified, and regulated in living organisms, while protein purification involves isolating one, or multiple, proteins from a complex mixture. Few proteins, however, are abundant in sufficient quantities to be isolated from their native hosts. Often, the researcher is tasked with figuring out ways to obtain an adequate concentration of the target molecule to facilitate protein expression and purification. After proteins are expressed sufficiently, they must be purified so that the desired protein(s) are separated from any other bioactive molecules within the cell sample.

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Magnetic IVD Coating for Disease Detection

In vitro Diagnostic (IVD) assays are used to diagnose infection or disease by specifically targeting a unique surface antigen or DNA sequence. IVD assays can be used to monitor a person’s overall health or to diagnose, treat, or prevent diseases. Magnetic beads have been developed to aid in molecular diagnostics, and provide the great advantage of simple manipulation via a magnet. In nucleic acid detection, magnetic beads act as a capture support for fast, easy, biomolecule separation. The surfaces of magnetic beads can be modified with IVD coatings for differing applications to allow for the specific identification and capture of target microbes. Magnetic IVD coatings can enhance the sensitivity and specificity of detection for the rapid diagnosis of a disease, point of care use, or for the quantitation of a specific microbe in a research setting.

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What are Magnetic Beads with Antigen and Antibody?

An antigen is a toxin or other foreign molecule that induces an immune response in the body. Antigens may exist in the environment in various forms but can also develop from inside the host. Antigens can present as proteins or sugars, and are located on the surfaces of all cells. When antigens invade the body, antibodies act as a first line of defense. They target and bind to antigens through what’s known as a paratope-epitope interaction (named after the binding regions on each molecule) and then undergo a series of steps to effectively eliminate them. Each specific antibody produced by the immune system is custom-fitted to the antigen that initially stimulated the immune response. The interaction of antibodies and antigens is so specific that it can be utilized in clinical diagnostics, specifically through the use of functionalized magnetic beads that can detect and capture a specific analyte.

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Latex Beads and Magnetic Latex Beads

Latex beads, also referred to as latex particles, are micrometer sized polystyrene beads made of polymer chains that form a spherical hydrophobic exterior. The beads bind proteins through passive adsorption and can be functionalized with chemical groups  to increase binding efficiency, for example towards amine groups on proteins. From here, the multivalent nature of antibodies allows them to bind to antigens on multiple latex beads simultaneously, causing the latex beads to agglutinate. 

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Superparamagnetic iron-oxide nanoparticles (SPION)-enhanced MRI imaging

Superparamagnetic iron-oxide nanoparticles (SPIONs) are small synthetic maghemite (γ-Fe2O3), magnetite (Fe3O4), hermatite (α-Fe2O3) particles and mixed oxides of iron with transition metal ions (e.g. copper, cobalt, nickel, and manganese) with a core from 10-100 nm in diameter. Magnetite and maghemite nanoparticles are the most widely used SPIONs in various biomedical applications both as diagnostic, therapeutic and imaging purposes.

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Cell Immunomagnetic separation protocol

Cell isolation is a critical step in the biomedical science and research sector and has been employed in in-vitro diagnostic tests and biological samples. The procedure of cell isolation lies on the specificity of a certain antibody toward its antigen. In recent decades magnetic beads have been implemented for the immunomagnetic separation protocols using both the superparamagnetic properties and antibody-antigen specificity which can be tuned to the experimental goal, and can produce high yields and highly enriched targets when used properly. Basically, cells of interest possess unique identifying surface antigens for the targeting beads. The immunomagnetic separation protocol follows these general steps:

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Silica magnetic beads

In recent decades, magnetic beads have been used in different sectors of biomedical science by introducing novel nanomaterials such as silica. Their size ranges from 1-5 um with a uniform disperse size distribution and sphere form. Magnetic Beads is highly suitable for automation since it requires no centrifugation, vacuum filtration procedures.

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Guide to the Magnetic Cell Separator

Cell separation is ubiquitously used in research and clinical settings where a target cell of interest needs to be isolated from a heterogenous mixture such as serum or plasma. It is used in several scientific disciplines such as immunology, where it helps identify cells present during immune responses, or in cancer research to help elucidate the tissue environment of tumors. Immunomagnetic cell separation uses monoclonal antibodies attached to magnetic beads to increase their ability to bind to target cells.In the magnetic cell separation system, a magnetic force collects the beads, allowing for targeted cell extraction at high yields.

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Magnetic beads DNA

Magnetic bead technology has been used for DNA isolation from biological samples with more advanced techniques in recent years due to the breakout of public health-threatening diseases requiring prompt diagnosis of the suspected disease. Isolation and amplification of the specific biomolecules is the downstream phase of numerous molecular methods such as detection, cloning, sequencing, amplification, hybridisation, cDNA synthesis, etc. and the presence of other cellular components and contaminating materials in the sample mixture makes this procedure very challenging.

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Magnetic Activated Cell Sorting

Magnetic Activated Cell Sorting (MACS) is a technique to enrich a specific cell type from a mixed population. Scientists or companies sort or isolate cells so they can study or grow colonies of a single type of cell. They may use that type of cell for a specific type of functional assay crucial to that cell type or they might be interested in stem cells. MACS technique emerged as a cheap alternative to cell sorting. MACS uses the highly specific antibody-antigen interaction to probe cells by their surface antigens by their specific antibody. Magnetic bead cell isolation has been implemented with MACS to offer a more precise cell sorting.

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Magnetic PCR Pathogen Detection

Regardless of the introduction of broad-spectrum antibiotics, pathogenic bacteria are still the main cause of life-threatening infectious diseases in the world. Prompt detection of pathogens with the lowest concentrations (<100 cfu/mL) without time-consuming procedures, such as culture or amplification by PCR is very important in disease diagnosis and the subsequent treatment regimen.

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Magnetic Gold Nanoparticles: Revolutionizing Diagnostic and Biomedical Applications

Introduction

With the advent of nanomedicine in recent decades numerous nanomaterials have been used for the formulation and synthesis of nanoparticles. A nanoparticle is defined as a tiny particle with a size ranging 1-100 nm. Among the different types of nanomaterials, magnetic gold nanoparticles (GNPs) have attracted much attention in the last decades. The two physical and chemical fundamental properties of GNPs are affected by their nanostructure – shape and crystal texture – which allows them to have numerous biomedical applications in prophylaxis, diagnosis and treatment.

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Magnetic Single Cell Isolation

As we have entered an age of personalized medicine, we have begun to understand that individual differences play a large role in disease expression and treatment options. This idea is also true of individual cells, the basic unit of life – a single cell is made of all of the necessary cell organelles that give that cell its functions and morphology. Therefore, while studying diseases’ phenotypes and developing new drugs, it is becoming increasingly important to study single cells instead of groups of cells.

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Magnetic beads antibody conjugation

Magnetic beads are used for biomagnetic separation procedures to enrich the population of a specific cell, or isolate biomolecules (such as protein, or nucleic acid) for purification purposes. Since the affinity between antibody and antigen is strong and specific, antibodies are often conjugated to the surface of magnetic beads – called “surface coating” – in order to bind their ligands for enrichment. 

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How a magnetic separation rack works

Magnetic separation has been an emerging technology in the recent decades in biomedical science and industry in which magnetic property and behavior – known as “magnetism” – of micro/nano-sized particles are employed for the separation of macromolecules of interest (e.g. nucleic acids, proteins, peptides etc) from biological samples or chemical suspensions.

Download our Free Guide on Biomagnetic Separation Scale-up HERE.
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Magnetic Nanoparticles: Preparation, Structure and Properties

Magnetic Nanoparticles (MNPs) are particles of nanosized range (10−9 nm)(usually ,100 nm in size) with unique properties of magnetic targeting, biocompatibility, surface modification characteristics and superparamagnetic properties. The application of magnetism in medical science was first introduced in the 1950s for “magnetic hyperthermia therapy” (cancer cell death) leading to various MNPs’ syntheses including Superparamagnetic iron oxide NPs (SPIONs).

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Magnetic Protein/peptide Purification Methods

With the advent of pharmaceutical biotechnologies in recent years proteins and peptides have been the main focus of numerous studies by researchers and companies. Peptide and proteins have various physiological functions in body (as hormones, enzyme substrates and inhibitors, biological regulators, structural components, signaling factors, catalyzers), peptide/protein-based drugs and biopharmaceuticals are a novel category of drugs, and any abnormality in their amino acid sequence or structural dysfunction can lead to severe diseases and pathological conditions (dwarfism, cystic fibrosis, thalassemia etc).

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Magnetic GST-fusion protein

A fusion protein is a protein composed of several domains (parts) that are encoded by separate genes and have been joined so that they are transcribed and translated as a single unit, producing a single polypeptide maintaining functional properties of each original protein. Fusion proteins can be created “in vivo” and “in vitro” by using recombinant DNA techniques for use in biological research or therapeutics. Fusion proteins occur naturally and commonly in cancer cells, where they may function as oncoproteins having different functions or physico-chemical patterns.

 

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Magnetic Protein Extraction

Proteomics is the study of the protein in an organism. Protein is a fundamental building block of life, and proteins are the workhorses within and between cells. Biochemical pathways are built out of enzymes and ligands—without them nothing would be accomplished; plants wouldn’t produce glucose, animals wouldn’t be able to digest food, the immune system would cease to exist, and all other biological processes would grind to a halt. The fundamental importance of proteins for life makes them an important topic of study. The first step in understanding protein structure and function is to extract them. Protein extraction is the process of isolating and purifying protein from samples of whole tissue, cell cultures, or biological fluids. The protein extraction protocol used is tailored to match the starting material and the end goals of the assay.

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Magnetic IVD assays for Malaria

In vitro diagnostics (IVD) are diagnostic tests performed on biological samples (e.g. blood, tissue) obtained from the body to detect DNA/RNA, microorganisms, or biological biomolecules (e.g. protein) associated with diseases, infections or medical conditions. The IVD tests are generally non-invasive, used both in professional healthcare settings and at home as rapid kits and are useful for early detection of diseases, prevent the spread of diseases and improve patient care and management.
     

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Magnetic Sandwich ELISA

The sandwich ELISA is one of the Enzyme-linked immunosorbent Assay (ELISA) methods which are analytical techniques for the detection of various compounds/analytes in a sample in the biomedical and research setting.

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Magnetic competitive ELISA

The Enzyme Linked Immunosorbent Assay (ELISA) is a gold standard analytical method for the detection of various compounds, and is the most commonly used immunoassay technique for labs and research.

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Magnetic cell sorting techniques

Isolation and identification of cell(s) is the prerequisite step for many fields of research, such as cell function and analysis, signaling and gene expression. Techniques that enable the rapid and accurate enrichment of target cell populations are therefore an area of substantial interest. The output of cell-sorting techniques from the cell suspension is based on higher efficacy or throughput, purity, and recovery. Based on the different principles used, the Cell sorting techniques are categorized into two general categories:

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Magnetic multiplex immunoassays

Immunoassay techniques are methods for the sensitive and specific quantitative detection of chemical substances such as hormones, drugs and specific proteins that utilize the highly specific binding between an antigen or hapten and homologous antibodies.

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Chemiluminescence vs Fluorescence

While chemiluminescence and fluorescence are used interchangeably, especially when referring to tracking strategies for magnetic separation in biosensors or in-vitro diagnostic assays, however, they are different concepts. They both give off a photon as an electron relaxes from a higher energy state to a lower energy state, but the difference lies in the method used to excite that electron to a higher energy state in the first place.  In fluorescence the electron is kicked up to a higher energy state by the addition of a photon. In chemiluminescence the electron is in a high-energy state due to the creation of an unstable intermediate in a chemical reaction. Light is released when the intermediate breaks down into the final products of the reaction.

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Background of Laemmli Buffer

The separation of charged biomolecules of a solution by their size and molecular weight is common for scientific studies and biomedical techniques for which various techniques of “electrophoresis” use an electric field, protein electrophoresis is among the widely used procedures.

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Magnetic beads protein A

Protein A

Protein A is a protein derived from the cell wall of the bacteria “Staphylococcus aureus” and can also be extracted from the culture supernatant. It has unique binding properties to the Fc and Fab region of IgG of some mammalian species (and to a lesser extent to IgM and IgA as well).

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Poly-L-lysine: what is it and what are its uses?

Poly-L-lysine is a synthetic amino acid polymer – a chain ofmultiple L-lysine amino acid monomers, with multiple industry and biomedical applications.

As a monomer, lysine contains two amino groups, called α-polylysine and ε-polylysine, based on which carbon they are on. Α-polylysine is composed of either L-lysine or D-lysine enantiomers, based on the chirality (“handedness”) of the lysine’s central carbon. Poly-L-lysine is a polypeptide formed from L-lysine monomers. Poly-D-lysine, a similar molecule used in similar applications, is a polymer formed with D-lysine units.

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Magnetic beads immunoprecipitation

Principles of immunoprecipitation with magnetic beads

Immunoprecipitation is one of the commonly used antibody-based techniques which relies on the specific affinity of an antibody to identify its target molecule within a biological sample for further analysis and research.

In a general protocol, antibody is pre-bound to a scaffold (magnetic beads or agarose beads) while it is incubated with lysates containing the target molecule. The mixture is then placed in the presence of a magnetic force. Using a classical magnetic rack the beads will be tightly held against the side of their container. The buffer simply can be removed from the container while the target molecule is still bound to the strongly immobilized beads to be finally eluted. This can also be done with free antibodies, which can be mixed with a cell lysate to bind their target, and then are bound to magnetic beads

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Sepmag is supporting the IVD industry to develop best in class Immunoassays

Magnetic separation is rapidly gaining popularity in the field of Life Sciences due to its numerous advantages. When considering the adoption of magnetic separation to purify biomaterial, it is crucial to recognize that the magnet itself plays an equally vital role as magnetic beads in achieving efficient biomagnetic separation.

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Magnetic Bead Fluorescent Immunoassays: How to use them in your research

Magnetic bead-based fluorescent immunoassays can detect and measure single or multiple analytes, such as certain proteins, present in one sample. The technology uses fluorescent magnetic beads, such as StrepTalontm or Luminex® beads, and detection antibodies to detect multiple analytes, and therefore answer multiple questions, simultaneously. So how do they work? 

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Cell Isolation

Importance and uses of cell isolation

Cells are the basic and fundamental units of organisms carrying genes for the biological function of that particular cell. Higher organisms (eukaryotes) are composed of 200 different types of cells (e.g. red blood cells, white blood cells, muscle cells, bone cells, nerve cells etc.) that have different gene expression profiles and even the same cell lines can present different genomes, transcriptomes and epigenomes during cell division and differentiation.

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What is transfection and what methods can be used to carry it out?

Transfection is a technique that makes it possible to modify the genetic content and therefore gene expression of host eukaryotic cells, both in vivo and in vitro. This makes transfection an important tool for drug discovery, the CRISPR-Cas9 technique, studying cell biology, cellular functions and molecular mechanisms of disease. The process centers on inserting proteins, nanoparticles or nucleic acids (such as cDNA, mRNA) into the cytoplasm of cells. 

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Conclusions to 'How to master your biomagnetic separation processes: consistency and monitoring'

Previously, the use of magnetic beads was limited to small volumes. The difficulty of scaling up beyond a few milliliters was misinterpreted as a limitation of the technology itself. However, as discussed in this e-book, the problem is not the biomagnetic separation process, but a lack of understanding of the physical processes governing it. Once you identify the key parameters that control the magnetic bead's behavior, it is easy to choose the right tools and methods to validate the process and replicate it at different volumes.

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Enzyme Immunoassay

Immunoassay tests are biochemical/bioanalytical methods that detect an “analyte” and quantify its concentration in a complex mixture of chemicals or biological fluids (e.g. serum or urine). Analytes can be a micro- or macromolecule (e.g. protein, nucleic acid, polysaccharide or lipid) or chemical substances (e.g. hormones, drugs).

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How monitoring your process can help you scale-up successfully

Once you have defined the required magnetic force, with a constant magnetic force separation device, it is simple to scale up production. Having validated the magnetic force at a small scale, the same force value can be used for a larger system, even in a different magnetic separation system. Because the conditions remain the same, efficiency (no losses) and batch consistency (no irreversible aggregation) are guaranteed.

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Magnetic separation rack

When designing a magnetic separation strategy, it is easy to get caught up in the properties of the superparamagnetic beads and how to coat them with the biomolecule of interest (antibodies, antigens, DNA, RNA, oligonucleotides, aptamers...). It is exciting to choose a bead and tailor its surface ligands to perfectly match your target molecule, but don’t stop there! The magnetic separation rack is equally important to a successful identification, isolation, or enrichment protocol. After all, a perfectly designed bead will be useless without a properly designed magnetic rack to efficiently recover it from the solution.

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Oligo dT-coated magnetic beads: the benefits of their application for mRNA purification

In the recent decades, proteins gained importance after the advent of more advanced analytical procedures and novel genetic or molecular engineering methods. Proteins are cell products and have various physiological functions in the body. Hence, any abnormality in gene expression (mRNA defects), amino acid sequence or structural dysfunction of proteins leads to severe diseases and pathological conditions.

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ELISA steps, principle, and how it works

The ELISA (Enzyme Linked ImmunoSorbent Assay) is the gold star immunoassay, used to detect the presence of a target protein. It is the standard procedure that all new assay technology is compared to during research and development. The ELISA is also fundamental to most clinical tests for diagnosing disease because it is highly sensitive, and is currently the best characterized and standardized method.

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Using biomagnetic separation curves to check re-suspension protocols

Under constant magnetic force conditions, optical monitoring of the biomagnetic separation process provides information on both when the separation is complete and the characteristics of the magnetic bead suspension.

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How to choose a protein purification system

Protein purification is a fundamental part of studying proteins, peptides, and nucleic acids, necessary for a wide range of clinical, research and industry applications. But choosing the most appropriate protein purification system can be challenging, especially for researchers who are just starting to think about automating their protein purification protocols.

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Parameterizing biomagnetic separation curves

Constant magnetic force separation systems generate the same conditions for all magnetic beads in the suspension. As bead behavior is consistent at every point of the working volume, any changes in the suspension's opacity can be directly related to changes in the suspension’s characteristics.

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How does magnetic bead separation work?

Magnetic bead separation is a quick, efficient, clean process that scientists use to replace filtration and centrifugation and separation techniques. Magnetic beads and particles are functionalized with antigens, antibodies, catalyzers, proteins or nucleic acids, enabling them to bind cells, bacteria, viruses and a wide array of other biological entities.
 

Magnetic bead separation enables complexes of magnetic beads and their bound materials to be separated from a complex mixture in solution with a single magnetic separation rack. The result is an isolated solution of your target biological elements which can be enriched and concentrated through this process. 

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Did you miss our invited talk at the Merck IVD Conference2023?

If you couldn’t attend our presentation ‘Developing protocols involving magnetic beads in manufacturing processes. What do we overlook?’, you have the opportunity to access it in Webinar format.

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Ensuring within-batch consistency

To ensure a consistent biomagnetic separation process, all the magnetic beads should experience the same conditions. Controlling the magnetic force is key to achieving consistency within and between batches, especially when scaling up. Classical magnetic separators generate a magnetic force that is very high on the side of the vessel closest to the magnet but declines rapidly with distance. The magnetic force experienced by beads in the retention area is therefore greatest, while the beads farthest away experience the lowest force.

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Protein assay

Proteins are one of the four macromolecule building blocks of life. The other three are carbohydrates, lipids, and nucleic acids. Proteins are long strings of amino acids that fold together into what are called “hierarchical structures” in order to perform specialized functions within the cells and tissues of all living organisms.

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Customizable NanoFrameworks

Customizable Nanoframeworks are one of the most exciting innovations in the world of nanochemistry. There are two main classifications of nanoframeworks. The first is the Metal-Organic framework (MOF). A MOG is a classification of a compound that consists of a metal linked to an organic ligand to form a coordinated structure in 1, 2 or 3 dimensions.

The second is a Covalent-Organic framework (COF), which is a crystalline porous organic framework with two or three dimensional properties. A COF is usually, but not always, limited to light elements (H, B, C, N and O) . Both possess a π-conjugated system and have a wide porous volume that can be tuned with the selection of a linker. This linker also has further effects on the electronic structure of the material. Thousands upon thousands of different, unique frameworks have been identified, leading to a variety of sizes that range from the nm to mm range. However, in all cases, the porosity of the framework benefits from a high surface area to volume ratio, leading to many different applications using a delivery mechanism that benefits from rapid diffusion. 

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GeneX, our distribution partner in India

Sepmag starts distributing its unique Biomagnetic separation systems in India, where Biotechnology industry is fast developing. In Vitro Diagnostic, protein purification and cell sorting companies working with magnetic beads will benefit enormously from the quality, reliability and monitoring that Sepmag technology offers to biotech enterprises around the globe.

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Ensuring batch-to-batch consistency – monitoring the separation process

Traditionally, biomagnetic separation users have not monitored the separation process. The nature of classical separators, where the magnetic force changes with the distance, means you can determine when the separation is complete (the buffer becomes transparent), but it is difficult to interpret the optical changes during the process. This is because every location sampled will have a different bead concentration due to their different speeds. In addition, it is difficult to compare different batches as even a small difference in the vessel’s position within the separator will affect the beads’ behavior.

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Cytiva’s patent application claims batch adsorption with magnetic beads is advantageous in the purification of sensitive plasma proteins

Batch adsorption with magnetic beads is considered a mild technique, which is advantageous for the purification of sensitive plasma proteins. Cytiva’s patent application shows excellent results using magnetic beads with suitable ligands for the purification of Factor VIII and von Willebrand Factor in solubilized cryoprecipitate, and Factor IX in cold supernatant. The magnetic separation system, with constant magnetic force, used in these experiments enables the straightforward transferring of the protocols to large-scale volumes.

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Don’t miss the next IVD Conference!

The Annual 2023 Two-Day IVD Conference will take place on March 21-22, 2023, in Rome, Italy.

SEPMAG will participate and provide helpful insights on how to develop protocols based on the utilization of magnetic beads in manufacturing processes.

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Immunoaffinity chromatography

Immunoaffinity chromatography is a method for separating target antibodies or antigens from a heterogeneous solution. It is column-based, which means that the solution is flowed through a column and eluted at the other end. The column is pre-functionalized with the capture antibody or antigen. The target protein is adsorbed onto the resin-bound capture protein and is retained in the column while the remaining solution is eluted. The fraction containing the target protein is later eluted and purified.

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Protein Purification using Magnetic Beads: One Protocol, All Sample Sizes

Cube Biotech’s video explores magnetic beads as a solution for large-scale protein purification using a Sepmag A400ml.

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What is RT-LAMP and what is it used for?

RT-LAMP is a Loop-Mediated Isothermal Amplification (LAMP) assay that uses a reverse transcription enzyme.

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Protein Extraction

Protein extraction is a key step for many proteomics research procedures, from ELISA to Western Blot. Proteins form the basis of all cells, tissue, and organisms. Proteins also initiate and mediate the thousands of biochemical pathways that govern an organism’s function. Biomedical studies of proteins can reveal information about pathways of disease, and the expression of the genetic code. But before proteins can be studied, they need to be extracted. Choosing the most appropriate protein extraction method is key to successful protein extraction.

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Immunomagnetic cell separation

Isolation and detection of a target molecule in cell therapeutics from a sample with high background debris or unwanted moleculesisa challenging task. Immunomagnetic-based separation is the most feasible technique to overcome the problems that come with the separation of cells and biomolecules from a complex matrix.

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Purification Techniques

The earliest chemists were on the hunt for new elements to add to the periodic table. Most of the chemistry that they were interested in doing was purification with the end goal of reaching a pure elemental substance. These chemists relied on a litany of methods—filtration, evaporation, distillation, and crystallization were some of the most used purification techniques for these discoveries. As the chemists were defining the elements, the biologists were trying to understand the human body, the cell, cellular organelles, and microbes. The point here is that in order to develop anything new we must first understand what everything is made of at the most basic and pure level. In modern science this means that we are trying to define matter beyond subatomic particles and we are attempting to map out every molecular pathway of disease. Our efforts to define complex systems by their purest constituents are rewarded by deep understanding and an ability to mimic, to engineer, develop, and create.

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Magnetic Nanoparticle Fluorescent Imaging

A fluorescent nanoparticle is a small particle containing a fluorophore that can be used to label biological material, such as a specific cell or tissue under fluorescence imaging. There are generally two locations for the particles to probe: ones that bind to the surface and others that bind internally. A large array of different nanoparticles can be used to achieve this, including but not limited to fluorescently doped silicas and sol-gels, hydrogels, hydrophobic organic polymers, and quantum dots. There are currently three main techniques for fluorescent problem.

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High molecular weight DNA: extraction using magnetic beads

Long read sequencing is making chromosome-scale assemblies, including diploid genomes, possible and is therefore improving our understanding of human genetic variation. But rapid improvements in long read sequencing capacity have been limited by the extraction of high molecular weight DNA. Magnetic bead-based high molecular weight DNA extraction limits DNA fragmentation, and is also less laborious and more cost-effective than other methods.

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Protein purification system

Proteins are large, complex biomolecules that perform a vast range of vital molecular functions in living organisms. Studies of the structure and function of proteins are helping to advance understanding of biology, but before proteins can be studied, they need to be isolated (i.e., purified). The best protein purification system for your application depends on the desired throughput, scale, and downstream application.

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Sepmag sponsors the UAB Nanoscience and Nanotechnology award (“Ciències Excel·leix"program).

The Faculty of Science celebrated the Ciències excel·leix Award ceremony on the last
December 16th. The aim of this program is to recognize and support outstanding first year students from the different Science degrees offered by the faculty.

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Scaling up – common problems

The variation in magnetic force with distance when using classical magnetic separators is rarely problematic at small volumes. The short distance between the farthest beads and the magnet means that even with the mild magnetic forces generated by a small permanent magnet, separation time is fast and the efficiency is high.

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Customizable Polymeric Bioparticles

With the recent innovations in nanofabrication, the use of a customizable bioparticle allows for many specified uses previously unimaginable for researchers. Highly sensitive purification, functional group modification or protection, and localized sensing are just a few specific applications that can be achieved using a customized bioparticle.

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Agarose Resin Chromatography

Using Agarose Resin chromatography allows for a versatile, prepacked column that enables small-scale, high-resolution size exclusion chromatography for preparation, characterization, and analysis of proteins and other biomolecules.

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What is the function of quenchers?

Quenchers are a substance that can decrease or end an effect or process. They are often used to absorb the excitation energy emitted by fluorophores (i.e., fluorescence), which signals that another process has taken place. Most re-emit much of the energy as visible light. Others, called dark quenchers, do not have native fluorescence and instead emit the energy as heat. Examples of common quenchers include iodide ions, molecular oxygen and acrylamide..

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Different types of Immunoassays

Immunoassays are tests that detect the presence of a specific molecule in a sample using antibody-antigen binding reactions. Antibodies bind to the specific structure of a particular antigen, making immunoassays highly specific: the antibody will only bind to a specific structure of a particular antigen. This makes antibodies effective reagents for detecting target molecules. As a result, immunoassays are a fundamental tool for hospitals, life science research and industry laboratories. Immunoassays come in a range of formats, and can be used to assess disease, track proteins, and detect environmental contamination.

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The industrial centrifuge

The industrial centrifuge plays an integral role in the production of more things than one would initially expect. It is a commonly used tool in the food and agricultural sector, At pharmaceutical and biotechnology companies, for environmental management, and in the chemical industry. The word industry conjures up images of combination and creation—adding materials together to produce a final product. However, the separation of materials is just as important as the combination of materials. We can't create a new product until we have pure reactants to work with. This is especially important in the pharmaceutical and biotechnological realms, where reactant purity is essential to the production of a product that is safe for human consumption. This is where the centrifuge comes in. The centrifuge is used to separate heterogeneous mixtures into components varying by density.

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DBCO click chemistry: what is it and what are its benefits?

DBCO click chemistry is a type of reaction that has underpinned recent advances in biomedical research, as well as pharmaceuticals and biotech. Copper(I)-free click chemistry, such as DBCO click reactions, enable fast and specific conjugation of target molecules under aqueous conditions and do not require toxic catalysts.

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What is poly dt?

Poly deoxythymine (poly dt) is a molecule made up of a string of deoxythymidines that are connected via 3' to 5' phosphodiester linkages. 

In other words, poly dt is a long chain of thymine nucleotides, which can be up to 20,000 nucleotides long. Also known as polythymidine, polythymidylic acid, and poly T, amongst other names, poly dt is a single-stranded sequence commonly used to purify messenger RNA (mRNA)..

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How to generate a constant magnetic force

A constant magnetic force across the whole working volume is key to consistency in biomagnetic separation processes. This ensures that all beads in the suspension experience the same force. Classical magnetic separators can't provide these conditions because the magnetic force they generate decreases with distance.

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Biodetection and biosensing: technology and applications

Biodetection and biosensors are widely used for diagnosing disease or infection, point-of-care monitoring and treatment, detecting toxins, environmental monitoring, forensics and research. Biosensing technology has a crucial role to play in future biomedicine and healthcare. Biodetection is a broad term that encompasses the global strategies in place for the detection of biological threats such as pathogens, infectious diseases, and biological weapons. In particular, portable biosensing instruments such as lab-on-chip technologies are opening up new possibilities for biodetection systems identifying outbreaks of infectious pathogens.

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Flow Cytometry Interpretation: how to know what your results mean

Flow cytometry interpretation can be challenging, particularly when conducting multiparametric analyses that typically create a large quantity of data points. Fluorescent activated cell sorting (FACS), a derivative of flow cytometry, has become an essential tool for modern life science and biomedical laboratories. It is one of two main methods for cell sorting, alongside magnetic bead separation. FACS is a method for rapidly analyzing cells based on multiple criteria simultaneously, and then physically sorting them based on this analysis. The results of the process are usually represented by dot plots: two or three-dimensional scatter plots, or sometimes a histogram. However, these plots can be tricky to interpret, so here are a few tips to help get you started.

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Why you can’t simply use a larger magnet for larger volumes

The key parameter for the biomagnetic separation processes is the magnetic force applied over the magnetic beads' suspension. The competition of this force with the drag force generated by the buffer viscosity will translate into the speed at which the magnetic beads separate.

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Cell Lysis Buffer

A cell lysis buffer is a critical first component to any isolation protocol. It is fundamental to the first step of protein or nucleic acid extraction as it aids in the chemical  breakdown of cell membranes and compartments, enabling target molecules to leave the cell. There are many types of lysis buffers; most are easy to make, but most are also commercially available. They are often included in kits for  immunoprecipitation, co-ip protocol, nucleic acid isolation, and others. When using a lysis buffer for protein capture the addition of protease inhibitors is generally recommended in order to protect proteins.

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Understanding magnetic force as the key parameter for biomagnetic separation processes

Biomagnetic separation has a wide range of applications in life sciences, from cell sorting to protein purification. But we regularly speak to laboratories and companies whose magnetic separation protocols lack necessary information on the key parameter: magnetic force.

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How to master your biomagnetic separation processes: consistency and monitoring

By improving the capture and isolation of biomolecules in complex matrices, magnetic beads have facilitated a leap forward in life science technologies..

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New e-book! How to master your biomagnetic separation processes

We are glad to announce the publication of a new e-book! Discover how you can take full advantage of this quick, efficient, and versatile technology.

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Meet with SEPMAG at AACC 2022!

We are excited to announce that we will be exhibiting at the 2022 AACC Clinical Lab Expo!

Visit our booth nr. 1559 to learn more on how to attain a safe and high-performing biomagnetic separation process. Discover how you can work with our systems with volumes from milliliters up to 50 liters, assuring batch consistency thanks to a constant magnetic force and an intuitive monitoring system.

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Good Manufacturing Practices

Good manufacturing practice, or GMP, is a set of standards that ensures that produced products meet a set of quality standards. Following GMP is crucial for the production of laboratory equipment, as it ensures that a manufactured product is able to meet predefined criteria. Most GMP practices follow the guidelines set by the FDA in the United States, and are promoted by the WHO.

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Nucleic Acid Isolation

Our understanding of genetic material has substantially increased since Friederich Miescher first extracted DNA in 1869. He discovered that a material exists within cells that precipitates out of acidic solution and dissolves into alkaline solution. He called it nuclein because it seemed to be located within the nucleus. It took until 1953 for the structure of DNA to be elucidated. It was during this time that procedures to isolate DNA began to emerge. Later, during the 1960's and 70's scientists were furiously untangling the cellular environment, and the discovery of RNA with its various forms and functions further refined DNA purification procedures.

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Hot gravity filtration and vacuum filtration in recrystallization

Recrystallization is the process of obtaining pure crystals from a compound containing impurities in a solvent. Hot gravity filtration is a process commonly used to remove these impurities from a solution prior to recrystallization. 

Hot filtration is a type of filtration where the filtration equipment and the sample are heated during the process. Hot filtration is needed for recrystallization when impurities exist in solution. Recrystallization requires a hot solution because the solution needs to be supersaturated for crystals to form on cooling. Hot solutions can hold more solute in suspension compared to a cold solution because the solubility of most solids increases with a rise in temperature. This means that a saturated solution will contain more dissolved solute if prepared at a higher temperature than at a cold temperature. When the hot solution then cools, it will be supersaturated – it will hold more dissolved solute than its cold equivalent would. 

The impurity may have a different solubility than the compound in certain solvents. The aim is to choose a solvent that dissolves the compound when heated, but that doesn’t dissolve the impurity at high temperatures. The impurity is then filtered out during the hot gravity filtration process.

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Nanobeads in biotechnology

Nanobeads have applications ranging from basic science research to clinical imaging and targeted drug delivery. Nanobeads are composites of nanoparticles. Nanoparticles are defined as being less than 100 nanometers in diameter, while nanobeads are usually around 50 to 200 nanometers in diameter. There are also microbeads, but these are much larger and have diameters of at least 1000 nanometers, or 1 micrometer, which is close to the size of a cell. Bacterial cell diameters range from 0.5 to 2 micrometers in diameter, and animal cells range from 10 to 30 micrometers in diameter. The size of nanobeads is very important to their function; partly because they are so much smaller than a cell, which enables them to be used for cell labeling and isolation. In the case of magnetic nanobeads, the nanometer size imparts the paramagnetic property that is so valuable for biomagnetic separation, clinical imaging (contrast enhanced magnetic resonance (MRI)), and therapeutics such as magnetic hyperthermia for targeted tumor destruction.

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Protein A Beads Optimization

Protein A beads like ELISA, Immunoprecipitation, antibody purification, and multiplex assays require the attachment of specific antibodies to a solid support such as a column, polystyrene plate, agarose bead, or superparamagnetic nanoparticle. There are a number of ways that antibodies are attached to solid supports. Some of these include:

  • covalently bonding the antibody’s primary amines directly to the surface
  • biotin-streptavidin affinity linkages
  • protein A and G
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Agarose Resin Chromatography

Introduction to Chemiluminescence immunoassays

Using Agarose Resin chromatography allows for a versatile, prepacked column that enables small-scale, high-resolution size exclusion chromatography for preparation, characterization, and analysis of proteins and other biomolecules. Size exclusion chromatography, also known as gel filtration, is a common technique used to separate compounds of small molecules, such as proteins, polysaccharides, and nucleic acids when in an aqueous solution. This can be extremely useful in numerous commercial applications, such as analysis and determination of an unknown sample, removal of large proteins for purification of a sample, for removing small molecules such as dyes and primers, and buffer exchanges. Such chromatography is purchased as a prepacked column that allows for increased resolution and analysis, quick runtime, with a high pH tolerance to allow applications to a variety of molecules and substances. One example of a commercially available separation is Superose 6 chromatography, found here.

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Magnetophoretic characterization of anisometric magnetic nanoparticles for hyperthermia

Introduction to Chemiluminescence immunoassays

One of the most promising applications of magnetic nanoparticles in medicine is their use for killing cancer cells by hyperthermia. You can place the nanoparticles in the right place by functionalizing them with antibodies specific to the membranes of the cancer cells and/or applying a magnetic field gradient to focalize them in a predetermined region of the body. If you apply an alternate magnetic field, the nanoparticles will heat and, they will kill the cells they are attached to. With the appropriate intensity and frequency, the increase in temperature will just be a few degrees and the effect would be very localized and selective.

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Chemiluminescent serological tests

Introduction to Chemiluminescence immunoassays

Serological tests are used to gain a deeper understanding of the immune response to pathogens and the tests help maintain community health by checking for antibodies in human biological samples. Chemiluminescence is a widely used system of reporting binding events. It is preferred because it uses a simple device for measurement, often one that measures output of visible light. This also allows the process to have a wide dynamic range, detecting light from binding events whether the sample is dilute or concentrated. Such detection is done with high sensitivity and with low background noise. The chemiluminescent magnetic microparticle immunoassay (CMIA) is a method developed to bring together the advantages of chemiluminescence and magnetic particles for immunoassays.

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Protein purification: the step-by-step process

Protein purification is the processes of isolating a protein of interest from its environment. In other words, from the other natural molecules surrounding the proteins in the natural niche in a host organism, or from a cell culture grown in a laboratory. Our protein purification handbook  explains that there are several available techniques and many options to consider, but the general procedure is the same. 

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Magnetic beads vs Agarose beads: Advantages of Magnetic Agarose in Protein Purification

Most current protein purification methods use agarose beads carrying affinity functionalities such as IMAC, Glutathione, or antibodies. The choice of these functional groups depends on the protein of interest to be purified, and a large variety is available, including pre-functionalized beads that can be coupled to biomolecules (see SEPMAG® protein purification handbook chapter 4 and 5).

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A general filtration process

Filtration is a simple technique used to separate solid particles from suspension in a liquid solution. There are many filtration methods available, but all are based on the same general principle: a heterogenous mixture is poured over a filter membrane. The filter membrane has pores of a particular size. Particles larger than the pores will be unable to pass through the membrane, while particles smaller than the pores will pass through unhindered. Additionally, all liquids will pass through. The final result of a filtration process is a collection of residue on the filtration membrane. This residue is therefore effectively separated from the rest of the mixture that passed through the membrane.

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Upstream processing

What is upstream and downstream processing?

Introduction to upstream and downstream processing. These terms are used more in the scientific industry, for example in pharmaceutical companies. Upstream is the first half of the process and everything associated with it. Downstream is the end of the biological process. Upstream processing being the first part of the biological process, it involves the growing of bacteria in media or culturing of cell lines. Companies use bacterial or human cells to harvest products of interest. There are many products that biological companies are interested in harvesting.

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Challenges with your Biomagnetic Separation process?

Visit Sepmag at AACR (booth 1456) and Experimental Biology (booth 1530)

SEPMAG is well known for helping IVD companies to improve, validate and scale up their biomagnetic separation processes. All this know-how on the physics behind the process also benefits researchers and industries in protein purification, cell sorting, and DNA/RNA capture.

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Immunoprecipitation with magnetic beads

Background on Immunoprecipitation

Immuno is a prefix that means you are talking about immunity. Immunity is how the body is protected against pathogens. The immune system has a system for recognizing foreign objects, then a system for combating the presence of the foreign object. For example in humans, T-cells are a type of immune cell that recognizes antigens, structures or molecules that are foriegn. Another important immune molecule is the antibody. Antibodies are shaped like the letter Y, and the two arms of the top of the Y recognize antigens. The specific part of an antigen that is recognized is called the epitope. The antibody recognizes the epitope by its structure and sequence of amino acids. This antibody-antigen interaction serves to help the body recognize antigens. When the interaction is strong enough, it also serves as a way to neutralize antigens. Another important aspect of antibodies is that they can have highly specific interactions with an epitope, and this interaction is strong as well, also known as a high affinity interaction. These two traits, the specificity and affinity, make antibodies a great tool as well! Let’s talk more about using antibodies for a particular tool, immunoprecipitation.


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Isolation of Cells

Techniques and investigations that require cell isolation

Cell isolation is a technique that is done in research labs and clinical settings. Cell isolation can be done in research settings to isolate a single cell to do research on it. There is a technique called patch clamp electrophysiology which measures voltage across a cell membrane. There are several ways to do this technique, either by inserting the pipette right into the membrane, or by taking a piece of the membrane off into the pipette so that molecules are still flowing through the membrane and the pipette which is connected to a device that can measure current. Another reason to isolate cells is to use them to study the effects of a drug on cell health. One grows cells in a dish in an optimized media for growth and stability. Then a drug can be introduced into the dish and one can observe how the cell physiology changes. The molecules released from the cells can also be studied or the change in the processes or proteins in the cell can also be studied with further purification or extraction techniques.


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