Bispecific rabbit Fab'-bovine serum albumin conjugate used in hemagglutination immunoassay for beta-microseminoprotein.
A polyclonal bispecific (bifunctional) antibody was prepared to develop a hemagglutination immunoassay for beta-microseminoprotein (beta-MSP), a predominant seminal protein. Three types of F(ab')2 fragments of rabbit IgG, affinity-purified anti-human red blood cell (RBC) F(ab')2 nonaffinity-purified anti-beta-MSP F(ab')2 and nonspecific (nonimmunized) F(ab')2, were mixed to obtain a F(ab')2 mixture containing 10% anti-RBC molecules and 10% anti-beta-MSP molecules. Fab' was obtained from the F(ab')2 mixture, and then reacted with maleimide-activated bovine serum albumin (BSA) at a molar ratio of 10:1. As estimated by the decrease in the maleimide content, approximately 7 Fab' molecules were introduced per one BSA molecule. The bispecific (anti-beta-MSP and anti-RBC) Fab'-BSA conjugate thus prepared was incubated successively with a human RBC suspension and with samples. In the presence of beta-MSP, RBCs become agglutinated, providing a test simple for forensic semen identification.
Yukawa
N
N
Department of Forensic Medicine, Tokai University School of Medicine, Kanagawa, Japan.
Osawa
M
M
Saito
T
T
Hasegawa
T
T
Matsuda
H
H
Takahama
K
K
Takeichi
S
S
D000017:ABO Blood-Group System / Q000276:immunology; D000818:Animals; D018033:Antibodies, Bispecific / Q000737:chemistry*; D000918:Antibody Specificity; D006386:Hemagglutination Tests / Q000379:methods*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D018796:Immunoconjugates / Q000737:chemistry*; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry*; D007074:Immunoglobulin G / Q000097:blood / Q000737:chemistry; D008297:Male; D029584:Prostatic Secretory Proteins; D011506:Proteins / Q000032:analysis* / Q000276:immunology*; D011817:Rabbits; D012661:Semen / Q000276:immunology*; D029589:Seminal Plasma Proteins; D012710:Serum Albumin, Bovine / Q000737:chemistry*
D000017:ABO Blood-Group System; D018033:Antibodies, Bispecific; D018796:Immunoconjugates; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D029584:Prostatic Secretory Proteins; D011506:Proteins; D029589:Seminal Plasma Proteins; C043076:beta-microseminoprotein; D012710:Serum Albumin, Bovine