[34] The amplified ELISA: Principles of and applications for the comparative quantitation of class and subclass antibodies and the distribution of antibodies and antigens in biochemical separates

摘要: Publisher Summary This chapter discusses one form of direct-binding enzyme immunoassay, the amplified enzyme-linked immunosorbent assay ELISA (a-ELISA), that has proved to be highly successful in measurement and quantitation antibodies different species isotypes a variety antigens. The a-ELISA differs from classical by addition several reaction steps use soluble, antibody–enzyme immune complex rather than chemically synthesized conjugate. A major technical advantage is absence any requirement for conjugation secondary antibody directly an enzyme. quantitated measuring amount colored product, p -nitrophenyl, accumulates. It found bovine antiserum human serum albumin (HSA) tested contains per milliliter 3.3 mg IgG 1 anti-HSA 2.5 2 anti-HSA, these data agree closely with values determined quantitative precipitation. determination levels cases where affinity, competition, antigen heterogeneity compromise collection also discussed chapter.