Murine monoclonal anti-H as typing reagent.
OBJECTIVE
The antigen H present on the surface of red cells in varying concentration, is maximum in O group red cells, but absent in Bombay phenotype individuals. This differentiation is generally detected by seed extracts of Ulex europaeus. The titre of such an extract is usually low and is subjected to batch variation. Hence, we carried out this study to raise potent murine monoclonal antibody against H antigen.


METHODS
Spleen cells of female BALB/c mice immunized with O group red cells were fused in presence of polyethylene glycol (PEG) 1500 with a mouse myeloma cell line Sp2/0 Ag14 in hypoxanthine aminopterine thymidine (HAT) selective medium and incubated at 37 degrees, 5 per cent CO(2) and 95 per cent humidity for a week.


RESULTS
The culture supernatants showing anti-H activity, were further subcloned and two clones 3E8A10 and 3E8A11 generated which showed a good potency, avidity and specificity.


CONCLUSIONS
The anti-H clones thus produced indigenously provided a potent reagent in distinguishing normal O group from Bombay phenotype individuals. The unlimited availability makes this reagent cost-effective to ensure a constant supply of hybrid clones with the similar specificities.
Iyer
Y S
YS
National Institute of Immunohaematology, ICMR, Mumbai, India. yegneswar2003@yahoo.com
Vasantha
K
K
Pujari
V
V
Jadhav
S
S
Ghosh
K
K
D000017:ABO Blood-Group System / Q000097:blood* / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002459:Cell Fusion; D045744:Cell Line, Tumor; D003470:Culture Media; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D013154:Spleen / Q000166:cytology
D000017:ABO Blood-Group System; D000911:Antibodies, Monoclonal; D003470:Culture Media