L4 - Lecture 4 ImmunoAssay E nzymeinked L I mmunoS orbent...

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Lecture 4 ImmunoAssay
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E L I S A nzyme- inked mmuno- orbent ssay Protein that facilitates a chemical reaction Immune system - antibodies a procedure for testing or measuring the activity of biochemical of a sample
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Enzyme-linked immunosorbent assay ( ELISA ) also known as an enzyme immunoassay ( EIA ) ; a biochemical technique: - used mainly in immunology to detect the presence of an antibody or an antigen in a sample; B cell
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Before the development of ELISA, the only method to perform immunoassay is r adio i mmuno a ssay ( RIA ). Either the antigen or the antibody is radioactively labelled (radioactive isotopes 125 I or 131 I ). The radioactivity provides the signal indicating whether a specific antigen or antibody is present in the samples (1960) Before ELISA
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Radioactive antigen Antibody with the radio active antigen Add unlabelled antigen The unlabelled antigen replaces those labelled antigen Measure those radioactivity of the labelled antigen Simplified version for RIA
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Radioactive antigen Antibody with the radio active antigen Add unlabelled antigen The unlabelled antigen replaces those labelled antigen Measure those radioactivity of the labelled antigen Simplified version for RIA Gamma Counter The main drawbacks: expensive hazardous when preparing and handling the radioactive antigen
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Y * * * * * * an unknown amount of antigen is introduced into a beaker a specific antibody is applied over the surface bind to the antigen another antibody linked to an enzyme is introduced another antibody linked to an enzyme will incorporate with the antibody A substrate is added to the system, the enzyme convert the substrate into a signal
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Antigens Antibodies Immunoassay ???
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Antigen • is a substance (or molecule) that triggers the production of an antibody (from the immune system when it is introduced into the body - is recognized as a foreign and potentially harmful invader • e.g. Pollen, virus, chemicals • The antibody will kill or neutralize the antigen
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L4 - Lecture 4 ImmunoAssay E nzymeinked L I mmunoS orbent...

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