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Preparation of specific antisera to 3 beta hydroxy 5 cholenoic acid by immunization with conjugates attached to protein through the carbon 3 position

, : Preparation of specific antisera to 3 beta hydroxy 5 cholenoic acid by immunization with conjugates attached to protein through the carbon 3 position. Chemical & Pharmaceutical Bulletin (Tokyo) 33(2): 831-836

3.beta.-Hydroxy-5-cholenoic acid-bovine serum albumin (BSA) conjugates were prepared by coupling of the haptens with the carrier protein through a 3-hemisuccinoyl or 3-hemiglutaroyl bridge. The antisera elicited by these new immunogens showed high affinity constants (Ka = 4.88 .times. 108 and 8.75 .times. 108/M) and excellent specificity to 3.beta.-hydroxy-5-cholenoic acid, exhibiting little or no cross-reactivities with cholesterol (0.001%) and related compounds. Though the antiserum obtained with the hemisuccinate-BSA conjugate was somewhat more specific than the other, they showed substantial the same reactivity, and should be useful for direct radioimmunoassay of 3.beta.-hydroxy-5-cholenoic acid in biological fluids without any chromatographic separation step prior to assay. [Oxidation of the cholesterol side chain is one of the more significant transformations in bile acid biosynthesis.].

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