Purification of blood-group substances in human hepatic bile (HHB) and immunological characterization of anti-HHB serum.
Blood-group substances (BGS) in human hepatic bile (HHB) of blood type A were purified 3657-7314-fold by ethanol sedimentation and column chromatography with DEAE Sephadex A-50 and Concanavalin A Sepharose 4B. An anti-HHB serum was raised by immunizing rabbits with the purified BGS in the HHB. Cross-reactions of the native anti-HHB serum with such human materials as type A blood cell, plasma, seminal fluid and saliva were eliminated by stepwise sequential absorptions with type A red blood cells, type O seminal fluid and type AB plasma. Precipitin titers of the native anti-HHB serum were x 320 and x 80 and those of the absorbed anti-HHB serum x 80 and x 20 against the immunogen and HHB, respectively. Organ and species specificity of the absorbed anti-HHB serum was checked by counter immunoelectrophoresis. It reacted specifically only with the immunogen and HHBs of various ABO blood types. No reactions with animal gallbladder biles were observed. In practical examinations, the specific anti-HHB serum identified 15 out of 20 (75%) HHB samples and 41 out of 46 (89%) human faeces samples. No reactions with animal gallbladder biles and animal faeces were observed.
Uchimura
Y
Y
Department of Forensic Medicine, Kumamoto University School of Medicine, Japan.
Furusawa
Y
Y
Hamada
C
C
Tsunenari
S
S
D000017:ABO Blood-Group System / Q000302:isolation & purification*; D000818:Animals; D000918:Antibody Specificity; D001646:Bile / Q000737:chemistry*; D001788:Blood Grouping and Crossmatching / Q000379:methods*; D001797:Blood Protein Electrophoresis; D002415:Cats; D002417:Cattle; D002645:Chickens; D004285:Dogs; D005243:Feces / Q000737:chemistry; D006801:Humans; D007106:Immune Sera; D005779:Immunodiffusion; D009928:Organ Specificity / Q000502:physiology; D013045:Species Specificity; D013552:Swine
D000017:ABO Blood-Group System; D007106:Immune Sera