Active sites antigen complex

Type III hypersensitivity is due to the presence of elevated levels of antigen-antibody complexes that deposit on basement membranes in tissues and vessels. Immune complex deposition activates complement to produce components with anaphylatoxic and chemotactic activities (C5a, C3a, C4a) that increase vascular permeability and recruit neutrophils to the site of complex deposition. Complex deposition and the action of lytic enzymes released by neutrophils can cause skin rashes, glomerulonephritis, and arthritis in these individuals. If patients have type III hypersensitivity against a particular antigen, clinical symptoms usually occur 3-4 days after exposure to the antigen. 

Subunits do not form a precipitate with antiphosphatase antibody however, there appear to be some antigenic determinates common to both subunits and active enzyme since subunits interfere with the precipitation of alkaline phosphatase. The alkaline phosphatase-antibody complex has 70% of the original enzymic activity as a suspension in solution. Therefore, the antibody does not bind to the active site of alkaline phosphatase, but it can still differentiate between monomers and dimers. 

The attachment of protein to the transducer surface for immunoassay is a difficult problem, since it must be achieved without interfering with the active site. If an unlabeled assay is to be performed it must also be such that subsequent non specific interaction with the surface can be inhibited. This requirement is often contrary to those for effective antibody-antigen complex formation. A surface close packed with antibody will be sterically hindered and its reaction with antigen inhibited. On the other hand, a suitably spaced packing allows non-specific interactions to occur and large false positive signals to be recorded. As mentioned earlier, this was demonstrated by Cullen and Lowe who used the surface plasmon resonance technique to probe specific and non-specific protein interactions at metal surfaces. (23). 

This leads to complete depolymerization and dissolution of the precipitate. In agreement with this mechanism, precipitation is not observed with monovalent antigens, nor with certain immunoglobulins which have become monovalent because of the intermolecular complexing of one of their active sites by an oligosaccharide sequence covalently linked to the polypeptide. 

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