N,N'-Bis(4-azidobenzoyl)cystine--a cleavable photoaffinity reagent

A water-soluble, cleavable, heterobifunctional photoaffinity label has been synthesized in one step from N-hydroxysuccinimidyl 4-azidobenzoate and cystine. The resultant compound, N,N'-bis(4-azidobenzoyl) cystine [(ABC)2], reacts with protein sulfhydryl groups through disulfide exchange to generate photoactive derivatives. Since [35S]cystine of high specific activity is readily available, it is possible to produce highly radioactive (ABC)2. ABC-derivatized ovalbumin is antigenic in vivo, and monoclonal antibodies specific for ABC have been produced. The antigen binding site of these antibodies was covalently labeled with ABC.