Screening and identification of stable transfectants of mouse soluble B lymphocyte stimulator.
- Author:
Chunhua FU
1
;
Ling TIAN
;
Yuquan WEI
;
Bing KAN
;
Jiong LI
;
Yanjun WEN
Author Information
1. Key Laboratory of Biotherapy of Human Diseases of Ministry of National Education and Cancer Center, West China Hospital, Sichuan University, Chengdu 610041, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Monoclonal;
B-Cell Activating Factor;
B-Lymphocytes;
immunology;
Cancer Vaccines;
biosynthesis;
genetics;
Cloning, Molecular;
Colonic Neoplasms;
pathology;
DNA, Complementary;
biosynthesis;
genetics;
Epitopes, B-Lymphocyte;
genetics;
Membrane Proteins;
biosynthesis;
genetics;
Mice;
Recombinant Proteins;
biosynthesis;
genetics;
Recombination, Genetic;
Transfection;
Tumor Cells, Cultured;
Tumor Necrosis Factor-alpha;
biosynthesis;
genetics
- From:
Journal of Biomedical Engineering
2004;21(6):897-900
- CountryChina
- Language:Chinese
-
Abstract:
Mouse colon cancer cells CT26 were transfected with constructed plasmid expressing mouse soluble B lymphocyte stimulator (msBlyS). Single cell clones were selected with 100 microg/ml Zeosin and subcloned by serial limiting dilution. Eight resistant transfectants were isolated and expanded, and five of them displayed the desirable msBlyS cDNA band amplified by semi-quantitative RT-PCR assay. Western blot analysis showed that only msBlyS molecules of the expected size were detected in the cell lysates from transfectants. The supernatant of transfectants could costimulate B cell proliferation in standard costimulation assay. Thus we have successfully screened the stable transfectants expressing high levels of msBlyS in CT26 cells, which could be used as cancer vaccines for further anti-tumor immunotherapy.
