The preparative refinement of steroid specific antibodies by affinity chromatography combined with electrophoretic desorption. Apparatus and method

The preparative refinement of antibodies to testosterone and 5α-dihydrotestosterone was achieved by prepurification (filtration, fractionation with ammoniumsulphate), followed by affinity chromatography of the antiserum. The affinity sorbent was synthesised by coupling 5α-dihydrotestosterone with Sepharose CL 6B. After elution of the interfering admixture, there was desorption by the use of an electric field, using a specially developed apparatus. This apparatus and the method employed provided preparative amounts of antibodies, without significant impairment of affinity. The interfering self fluorescence of the serum was greatly diminished by this refinement procedure. The refined antibodies obtained were suitable for use with sensitive fluorometric testings (i.e. homogeneous fluoroimmunoassay).