Enzyme-linked immunosorbent assays antibody inhibition

Antiphospholipid antibodies include lupus anticoagulants (LAs) and anticardi-olipin (aCL) antibodies. Lupus anticoagulants are immunoglobulins that are characterized by their ability to inhibit phospholipid-dependent coagulation assays. In contrast, aCL antibodies are measured in an enzyme-linked immunosorbent assay... 

Enzyme-Linked Immunosorbent Assays (ELISA). Three methods are commonly used direct competition, double antibody sandwish and antibody inhibition. 

Figure 1. Dose-response antigen inhibition curves derived from ELISA (enzyme-linked immunosorbant assay), utilizing an antiserum raised against synthetic pigment-dispersing factor (PDF) of Romalea, Assay conditions antigen coat, 200 fmol Romalea PDF/well primary antibody dilution 1 140,000 secondary antibody (peroxidase-labelled antirabbit IgG), 1 1000 other details as described earlier (63). When no Romalea PDF is present in the assay, the obtained value is defined as 100%.

The most common types of assays employed to quantitate protein concentrations in biological matrices are listed in Table 32.4. Enzyme-linked immunosorbent assays (ELISAs), radioimmunoassays (RIAs), and immunoradiometric assays (IRMAs) require protein-specific antibodies, labeled proteins, or labeled antibodies as reagents, and are generally competitive inhibition assays. Radioimmunoassays measure concentrations by displacing ligands from cell-bound receptors. The most common assay, the... 

To assess the activity of a-gal-mannose glycoconjugates binding to the anti-Gal antibodies, a previously reported inhibition-type enzyme-linked immunosorbent assay (ELISA) was conducted [21]. Mouse laminin having a-gal epitopes were fixed on ELISA plate as solid phase antigens. Test glycoconjugates were then incubated with human anti-Gal antibodies on the ELISA plate. The plate was washed and incubated with horseradish peroxidase (HRP) conjugated anti-human IgG antibody. 

The evaluation of the binding affinities of synthesized a-Gal-containing polymers 7 to anti-Gal antibodies was accomplished by inhibition ELISA (enzyme-linked immunosorbent assay) with purified human (male, blood type AB) anti-Gal antibody as the primary antibody and mouse laminin as a natural source of a-Gal. The concentrations of a-Gal polymers at 50% inhibition (IC50) of anti-Gal antibody binding to a-Gal epitopes on mouse laminin were measured and summarized in Table 1. 

The procedures for production of specific antibodies and their application in a competitive inhibition ELISA (Enzyme-Linked Immunosorbent Assay) are discussed in detail in the preceding chapter (Vanderlaan et al., this volume). In addition, other comprehensive overviews of the immunoassay development process in the pesticide field are available in general, a... 

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