Chemiluminescent Immunoassay
September 02, 2021
What is Chemiluminescent Immunoassay Chemiluminescent immunoassay (CLIA) is an immunoassay technique where the label, i.e. the true “indicator” of the analytic reaction, is a luminescent molecule. In general, luminescence is the emission of visible or near-visible radiation which is generated when an electron transition from an excited state to a ground state. The resultant potential energy in the atom gets released in the form of light. (Cinquanta, Fontana and Bizzaro, 2017) Chemiluminescent immunoassays offer one of the best solutions for the quantification of low concentrations of specific analytes from a complex mixture for in vitro diagnostic industry. They can also be used as diagnostic tools in medicine, as well as being used in several other different industries for various applications. (Lorne Laboratories, 2021) How Does Chemiluminescence Work Chemiluminescence immunoassay (CLIA) is an assay that combines the chemiluminescence techniques with immunochemical reactions. The assay format is similar to enzyme-linked immunoassays, usually based on heterogeneous assays where antibodies or antigens are immobilized on a solid phase but one of the components is conjugated with a chemiluminescent label. (Lorne Laboratories, 2021) (BrightTALK, 2021) CLIA utilizes chemical probes which could generate light emission through a chemical reactions to label the antibody. (Creative-diagnostics, 2021) The Benefits of Chemiluminescence Immunoassay In recent years, CLIA has gained increasing attention in different fields, including life science, clinical diagnosis, environmental monitoring, food safety, and pharmaceutical analysis because there are benefits of using chemiluminescence such as: ultra-sensitive good specificity wide range of applications simple equipment wide linear range easier in operation and design can detect small amounts of the biological molecule does not require long incubations and the addition of stopping reagents (Creative-diagnostics, 2021) Key Types of Chemiluminescent Immunoassay Methods Chemiluminescent methods that have been commonly seen are direct - using luminophore markers, indirect—using enzyme markers, or electrogenerated – typically nanomaterials based. (Cinquanta, Fontana and Bizzaro, 2017) (Ju, Lai and Yan, 2017) Indirect CLIA, using enzyme markers This common CLIA method involves an enzyme, typically horseradish peroxidase (HRP) or alkaline phosphatase (AP), conjugated to a secondary antibody. This HRP– or AP–secondary antibody complex then catalyzes the conversion of chemiluminescent substrate, luminol and CSPD reagent respectively. These substrates, when oxidized, provide an easy means of signal detection and quantification due to prolonged signal emission. (Shimadzu, 2021) Direct CLIA, using luminophore markers This method commonly uses acridinium esters as luminophores conjugated directly to the primary monoclonal antibody to generate a signal. Acridinium esters are highly specific labels that spontaneously oxidiz...
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