The preparation and use of antibodies as diagnostic biochemical probes

This chapter discusses various techniques in which antibodies to specific proteins or small molecules are used as analytical biochemical tools and with some of the procedures used to elicit and purify these antibody preparations to the level of purity required. It has become more obvious that the highly specific binding of antibodies to their respective antigens can be an invaluable property to utilize in purification, determination of functional role, tissue and subcellular localization, and identification and quantitation of enzymes or small molecules (metabolites, hormones, etc). In particular, a recent study has been conducted on the refolding of bovine serum albumin catalyzed by either glutathione or rat liver disulfide interchange enzyme, and various antigenically active fragments that represent domains and subdomains of the molecule have been isolated by Teale and Benjamin. Restricted populations of antibody to the albumin molecule were obtained by fractionating the antisera to the native, intact molecule on immunoabsorbents bearing these fragments. The chapter also discusses the antibodies used as probes of conformational state of a single protein.