Anti-bilirubin monoclonal antibody. III. Preparation and properties of monoclonal antibodies to unconjugated bilirubin-IX alpha.
Anti-bilirubin-IX alpha monoclonal antibodies exclusively specific for unconjugated bilirubin-IX alpha are prepared and characterized. Using modified MBS (metamaleimidobenzoyl-N-hydroxysuccinimide ester) method, bilirubin-IX alpha was covalently coupled to bovine serum albumin (BSA) retaining its intramolecular hydrogen bonds as well as three-dimensional structure. Somatic cell fusion was performed between murine spleen cells immunized with the bilirubin-IX alpha-BSA and murine myeloma P3-X63-Ag8-U1 cells. After screening assay, 58 clones were identified which were producing antibodies not to albumin nor MBS, but to haptenic bilirubin-IX alpha. In inhibition analysis, the antibodies in the culture supernatant reacted only with bilirubin-IX alpha to a varying degree, but did not react with conjugated bilirubin-IX alpha, including bilirubin-IX alpha monoglucuronide, bilirubin-IX alpha diglucuronide, and ditauronic bilirubin-IX alpha.
Okamura
Y
Y
Department of Biochemical Genetics, Tokyo Medical and Dental University, Japan.
Yamazaki
M
M
Yamaguchi
T
T
Komoda
Y
Y
Sugimoto
A
A
Nakajima
H
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000918:Antibody Specificity; D001663:Bilirubin / Q000276:immunology*; D002459:Cell Fusion; D003432:Cross-Linking Reagents; D005260:Female; D006825:Hybridomas / Q000276:immunology; D007536:Isomerism; D051379:Mice; D008807:Mice, Inbred BALB C; D012710:Serum Albumin, Bovine / Q000737:chemistry; D013388:Succinimides
D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D013388:Succinimides; D012710:Serum Albumin, Bovine; C013097:3-maleimidobenzoyl N-hydroxysuccinimide; C028529:photobilirubin; D001663:Bilirubin