[Preparation of monoclonal antibodies against multiple antigens].
OBJECTIVE
To prepare monoclonal antibodies (mAbs) against multiple antigens by single cell fusion.


METHODS
BALB/c mice were immunized with the multiple antigens, namely alpha fetoprotein (AFP), carcinoembryonic antigen (CEA), HBsAgiHBcAg and HBeAg, and hybridomas were employed using PEG as the fusing agent. The hybridoma cells were respectively screened with AFP, CEA, HBsAg, HBcAg and HBeAg by enzyme-linked immunosorbent assay and limited dilution. The mAbs were purified by protein G affinity chromatography, its subtype was identified, the affinity constants (K(a)) were determined and the specificity was analyzed by Western blotting.


RESULTS
Twenty hybridoma cell lines were obtained by single cell fusion, including 5 cell lines against AFP, 6 against CEA, 3 against HBsAg, 4 against HBcAg, and 2 against HBeAg. The subtypes of some hybridoma cell lines positive for the mAbs were identified as the immunoglobulin G1 (IgG1), with K(a) ranging from 1x10(9) M(-1) to x10(11) M(-1). Western blot analysis showed that all the mAbs strongly and specifically bound to their respective antigens.


CONCLUSIONS
The mAbs against multiple antigens have been obtained by single cell-fusion, which increases the production of mAbs and reduces the time of preparation.
Peng
Dan-dan
DD
Institute of Tropical Medicine, Southern Medical University, Guangzhou 510515, China. dannypen@163.com
Ning
Yun-shan
YS
Li
Yan
Y
Hong
Yan-hua
YH
Wang
Yun-dan
YD
Wu
Fen-fang
FF
Li
Ming
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D002272:Carcinoembryonic Antigen / Q000276:immunology*; D002459:Cell Fusion / Q000379:methods; D005260:Female; D006511:Hepatitis B Antigens / Q000276:immunology*; D006514:Hepatitis B Surface Antigens / Q000276:immunology; D006513:Hepatitis B e Antigens / Q000276:immunology; D006801:Humans; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D000509:alpha-Fetoproteins / Q000276:immunology*
D000911:Antibodies, Monoclonal; D002272:Carcinoembryonic Antigen; D006511:Hepatitis B Antigens; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; D000509:alpha-Fetoproteins