Preparation and comparative properties of antisera against glyco-3 beta,12 alpha-dihydroxy-5-cholen-24-oic acid linked to bovine serum albumin at the C-3, C-15 alpha, and C-24 positions.
Anti glyco-3 beta,12 alpha-dihydroxy-5-cholen-24-oic acid antisera were prepared by immunizing rabbits with hapten-bovine serum albumin (BSA) conjugates coupled at the C-3, C-15 alpha, and C-24 positions on the bile acid molecule, and their properties were investigated by heterologous combination assay using 125I-labeled tracer. The antiserum raised against the C-3 BSA conjugate showed poor titer and specificity, while the antisera from the other two conjugates showed satisfactorily high affinity constants (Ka = 5.0 x 10(8) and 7.0 x 10(8) M-1) and reasonable specificity, exhibiting negligible cross-reactivities with other major human bile acids and cholesterol. Among the unsaturated bile acids tested, high reactivity was observed with tauro-3 beta,12 alpha-dihydroxy-5-cholen-24-oic acid, which suggested that bridge phenomena were significant in this assay system.
Ikegawa
S
S
Kinoshita
J
J
Takeshita
K
K
Sato
H
H
Tohma
M
M
D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification*; D002793:Cholic Acids / Q000276:immunology*; D003429:Cross Reactions; D007457:Iodine Radioisotopes; D011485:Protein Binding; D012710:Serum Albumin, Bovine / Q000276:immunology
D000906:Antibodies; D002793:Cholic Acids; D007457:Iodine Radioisotopes; C062129:glyco-3,12-dihydroxy-5-cholen-24-oic acid; D012710:Serum Albumin, Bovine