Preparation of monoclonal antibody based on single intrasplenic immunization of plasmid DNA.
- Author:
Hong ZHENG
1
;
Shuang-Yan HAN
;
Ying LIN
;
Xiao-Ning WANG
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies, Monoclonal; biosynthesis; genetics; immunology; Antibody Specificity; DNA; genetics; immunology; Female; Gene Fusion; Immunization; Interleukin-2; genetics; immunology; Mice; Mice, Inbred BALB C; Plasmids; genetics; Profilins; genetics; immunology; Spleen; immunology; metabolism
- From: Chinese Journal of Biotechnology 2007;23(4):710-714
- CountryChina
- Language:Chinese
-
Abstract:
UNLABELLEDTo set up a new rapid and efficient way for the preparation of specific monoclonal antibodies (MAbs) with plasmid DNA immunization.
METHODSThe fusion gene of IL-2 signal peptide and profilin 1 by 'overlapping PCR' was obtained and inserted into the vector pVAX1 to construct the recombinant plasmid pVAX-IL2-profl. Another fusion gene of IgG kappa chain signal peptide and profilin 1 by 'no template PCR' was obtained and inserted into the vector pVAX1 to construct the recombinant plasmid pVAX-Igkappa-prof1. BALB/c mice were single intrasplenic immunized with plasmid DNA. Results After cell fusion and hybridomas screening by indirect ELISA, we charactered two mAbs (1D8A2B4 and 4B12A5E3) against profilin 1. The MAbs isotype were determined as IgM and IgG3.
CONCLUSIONA single intrasplenic plasmid DNA immunization is rapid and efficient and can be used as a helpful tool for the production of specific mAb.
