METHOD OF DETERMINATION OF AUTOANTIBODY LEVEL BY MEANS OF ENZYME IMMUNOASSAY

摘要

The method for quantitative determination of the level of natural autoantibodies in human biological fluids, when as a solid phase of physical sorption is used the solid phase of physical sorption, coated with streptavidin, and the solid phase of physical sorption is treated with preliminary biotinylated antigen and blocking agent for closing the sites of nonspecific binding at the solid phase of physical sorption, for which purpose are used proteins, biotinylated according to standard procedure. As the conjugate-containing solution are used enzyme-labeled monoclonal and polyclonal antibodies, which react with one or all isotypes of human immunoglobulins. In addition, the tested biological fluid is preliminary diluted in a buffer, containing proteins which are used for closing the sites of nonspecific binding at solid phase of physical sorption, and also substances protecting natural autoantibodies from destruction during heat treatment, and subjected to heat treatment. For each tested specimen of biological fluid, a control solid phase of physical sorption is used, and the number of natural autoantibodies is determined with the use of a calibration curve which is plotted using monoclonal or polyclonal antibodies to antigen.