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Process Validation Protocol

Good manufacturing practice is an essential part of the production of human drugs, veterinary drugs, biological and biotechnology products, and pharmaceutical ingredients. These commercial processes are subject to regulatory oversight and must ensure that every aspect of the production process is carefully scrutinized. The purpose of process validation is to collect data and scientifically analyze the production process from conception to large scale production. An updated process validation protocol is essential to ensuring product quality and consistency. Many laws have been established to mandate process validation in order to protect consumers, especially in the case of pharmaceutical products.

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A magnetic separator for biological application

Magnetic beads and separators have grown in popularity as a biotechnological tool over the past few decades. As we gained experience with the technology, we learned that all magnetic separators are not created equal. It turns out that it isn’t just the magnitude of the magnetic force that improves separation time, it is the way it spreads out over the working area that matters the most. A homogeneous magnetic separator is designed to ensure that every point within the working volume experiences the same magnetic force. This is especially important when scaling up a biomagnetic separation process from milliliters to liters.

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Formalin-fixed paraffin-embedded (ffpe) and frozen tissue preservation

The collection of tissue is a fundamental procedure for research and clinical biology. Before collection it is important to consider which method will be used to preserve the tissue and prepare it for histology or molecular analysis. There are two options to choose from when collecting and preserving tissue: frozen or formalin-fixed paraffin-embedded (ffpe). Each has its strengths and weaknesses, but these are only apparent when the intended use of the tissue is considered.

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Advanced molecular diagnostics

Molecular diagnostics entails the analysis of biomarkers to help diagnose, track the progression or, or determine risk factors and prognosis of disease. Biomarkers have been identified within the realm of genomics, epigenomics, transcriptomics, proetomics, metabolomics, and lipidomics:

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Magnetic activated cell sorting: technology and advantages

 

The objective of magnetic bead cell isolation is to enrich a specific cell type from a mixed population. The technique is also be used to isolate proteins, DNA, and RNA for further research or therapeutic purposes. The versatility and specificity of magnetic bead cell isolation is made possible by functionalized bead surfaces that specifically recognize a target molecule. Magnetic beads are composed of a ferrous iron-oxide core surrounded by a polymer shell, or a magnetic ‘pigment’ embedded in a polymer matrix.

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Anti Human IgG

Anti human IgG is the most prevalent antibody isotype in human serum. It plays a critical role in the adaptive immune response. When a pathogen invades a human body it is intercepted by a white blood cell called a B-cell. These B-cell lymphocytes have cell surface receptors that bind a wide array of pathogens. Once activated, the B cell divides and produces offspring cells which secrete very specific antibodies capable of identifying that exact invading pathogen. These antibodies recognize a specific antigen binding site.

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GST fusion protein

A GST (Glutathione-S-transferase) fusion protein is useful for affinity chromatography and immunoprecipitation. The natural form of GST is an enzyme that catalyzes the protective mechanisms of glutathione. Glutathione is an antioxidant that prevents cell damage by reactive oxygen species. However, the GST fusion protein is not natural. It is a genetically engineered protein that has become a useful biotechnological tool.

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Protein A beads

Immunoprecipitation, ELISA, 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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Biotinylation Kit

Biotinylation means attaching a biotin tag to a molecule. The purpose of biotinylation is to create a controlled site for biotin-streptavidin affinity binding. Biotin and streptavidin have an extraordinary affinity for each other (Kd=10^-15). This affinity is resistant to changes in temperature, pH, and salt concentration and is extremely specific. These properties make biotinylation a useful tool for engineers who are developing new purification and detection methods. A commercially available biotinylation kit  makes the process even easier.

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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, centrifugation and separation techniques. Magnetic beads and particles are used as carriers of antigens, antibodies, catalyzers, proteins and nucleic acids, enabling action on cells, bacteria, viruses and other biological entities.
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