[Establishment of hybridoma cell lines producing monoclonal antibodies against-p30 antigen].
BALB/c mice were immunized with the purified p30 antigen. SP2/0 myeloma cells and immune spleen cells were fused with 50% PEG (Sigma, MW 3,350-4,000). The cell fusion rate was 93.95%, and the antibody producing rate 21.01%. The technique of limiting dilution was used for cloning of the hybridoma cells. Two hybridoma cell lines E8 and G1 secreting McAb against p30 antigen were obtained. The number of chromosome of both E8 and G1 cell lines was 98.7 +/- 6.54 and 97.7 +/- 7.77, respectively. Results of the PAGE of the ascites generated by the hybridoma cell lines E8 and G1 showed a thick protein band at the gamma-region which was absent from the ascites generated by the SP 2/0 myeloma cells. The immunoglobulin of the McAb E8 and G1 belonged to IgG2 subclass. The results of the immunohistochemical method using the horseradish peroxidase conjugated antibody showed that both E8 and G1 McAb only reacted with epithelial cells of the normal human prostatic glands and their ducts, but did not cross-react with other thirty-four different kinds of normal human tissues. The results of inhibiting ELISA test showed that both E8 and G1 McAb were only inhibited by the human seminal plasma and p30, weakly inhibited by the adult male's urine, but were not inhibited by other eight different kinds of human body fluids and secretions as well as semen from eight different species of animals. It was concluded that McAb of E8 and G1 were organ and species specific.(ABSTRACT TRUNCATED AT 250 WORDS)
Li
Y
Y
Wu
M
M
Zhang
L
L
Zhao
Q
Q
Zeng
Y
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D002460:Cell Line; D006801:Humans; D006825:Hybridomas / Q000378:metabolism*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D029584:Prostatic Secretory Proteins; D011506:Proteins / Q000276:immunology*; D012661:Semen / Q000276:immunology; D029589:Seminal Plasma Proteins; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D029584:Prostatic Secretory Proteins; D011506:Proteins; D029589:Seminal Plasma Proteins; C043076:beta-microseminoprotein