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Dr. Anabel Lermo

Dr. Anabel Lermo
Dr. Anabel Lermo is a Research Scientist within the field of In-Vitro Diag- nostic (IVD). She holds a PhD in Chemistry from the UAB of Barcelo- na. After her postdoctoral work in the field of immunoassay development applied to biosensor design at the Analytical Chemistry Division UAB, she joined Biokit (a company of the Werfen group) for 7 years, developing chemiluminescent immunoassays for automated platforms in the field of infectious diseases. Later she worked for Pragmatic Diagnostics develo- ping new assays where immunometric technologies are integrated with appropriate analytical platforms for IVD industry.

Recent Posts

 

Basic Guide for optimizing Chemiluminescent immunoassay (CLIA) performance and scaling-up

There are a few more considerations for optimizing the CLIA assay that in this chapter will be discussed. The following considerations are related to the performance of your assay.

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Chemiluminescent immunoassay (CLIA) Tracer Optimization

The tracer, the antigen or antibody labelled with a chemiluminescent tag for CLIA, is the next vital optimization step of a chemiluminescent immunoassay. As mentioned earlier, chemiluminescent labels generate light from a chemical reaction. Widely used CLIA labels  are based on luminol derivatives or acridinium esters.

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Most common Chemiluminescent immunoassay (CLIA) formats

There are several types of CLIA formats that can be used depending on the target analyte of your assay. The choice of assay format will impact four major aspects of development. The first will be the choice of magnetic bead coated with antigen/s or antibody/ies for binding the  target analyte. The tracer will then be required to match the target analyte using  a conjugated antibody/ies or antigen/s conjugatedith a CLIA label. The assay buffer will need to be optimized to improve the specificity and sensitivity for each step. Lastly there will be components such as blockers, other linking molecules or stabilizing molecules. These aspects can be optimized once an assay format is chosen. 

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Basic guide for successful magnetic bead conjugation in Chemiluminescent immunoassay (CLIA)

For a successful procedure for the magnetic bead conjugation, there are three important aspects to take in consideration when designing the assay: i) the planning of the conjugation protocol, ii) the density of the functional groups on the surface of the magnetic beads, and iii) the controlled magnetic separation of the beads.

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Aspects of available Chemiluminescent immunoassay (CLIA) kits in the market

When developing a CLIA it is helpful to understand what is available commercially. CLIA kits are available from many different companies that formulate  reagents or components of the assay specifics for the analyte to detect and tailor made for the company analyser (platform). The solid phase can be based on superparamagnetic beads or polystyrene beads usually kept in liquid formulation. In both cases these beads require a CLIA-label reagent, discussed in more detail earlier in this ebook.

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Preliminary considerations for Chemiluminescent immunoassay (CLIA) design

Designing CLIA assays requires the consideration of different aspects, encompassing the raw materials for the reagent development & methods selection, together with the choice of the assay format. Material suppliers are a key factor for a successful design and development of an assay. An ideal supplier should be able to provide required raw materials not only at a reliable cost but also available to provide the required bulk quantities for scaling up the reagent. Moreover, suppliers should provide different lots to assess the lot to lot variability to check the impact in the assay to be developed.

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Limitations and perspectives of Chemiluminescent immunoassay (CLIA)

Advantages of the CLIA

There are many types of assays that can be performed for detection of a molecule of interest, all with their own advantages and disadvantages. Many scientists choose to perform chemiluminescent immunoassays over the enzyme-linked immunosorbent assays (ELISA), fluorescence or radioimmunoassays. This is because the CLIA has been shown to have an improve detection at lower concentration and a wide dynamic range.

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CLIA-label protein conjugation

Once an optimal CLIA label (also called CLIA tag) is chosen, it must be conjugated to the protein which will bind to the analyte  desired to detect. Isoluminol or Acridinium ester derivatives are often used as CLIA tags. 

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Magnetic beads in Chemiluminescent immunoassay (CLIA)

How can magnetic beads improve CLIA tests?

The combination of CLIA and magnetic beads brings together all the advantages of both parts. CLIA is known for its high sensitivity which allows the detection of analytes at very low concentrations, and thus providing an excellent limits of detection in a wide dynamic range.

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Overview on Chemiluminescent immunoassay (CLIA)

General Introduction

Luminescence is the emission of light, and it can occur in many ways. In research and biomedical industry fluorescence and chemiluminescence are often used. Fluorescence is when light is absorbed then emitted by a substance. A photon of a higher energy state is absorbed, then a lower energy photon is emitted in another range of the electromagnetic spectrum. 

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