Expression, identification and bioactivity characterization of an anti-CD3/anti-CD20 bispecific single-chain antibody / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 289-293, 2005.
Article
in Chinese
| WPRIM
| ID: wpr-305283
ABSTRACT
The synthetic gene with 1640bp encoding for the anti-CD3/anti-CD20 bispecific single-chain antibody was designed and obtained by SOE (splicing by overlap extension) PCR. The cDNA was cloned into Flp-In expression vector pcDNA5/FRT and transfeced into Flp-In CHO cells to generate a stable expression cell line with a capacity for expressing anti-CD3/anti-CD20 bispecific single-chain antibody at 300 microg/L. The protein, which had a molecular weight of about 70 kD,was purified by Ni-NTA affinity chromatography and identified by SDS-PAGE and Western-blot analysis. Immunofluorescence assay and cellular rosetting showed that it can react specifically on Jurkat (CD3+) and Ramous (CD20+) cells. The lysis of human PBL against CD20-positive lymphoma Ramous cells in the presence of the anti-CD3/anti-CD20 bispecific single-chain antibody can observed by microscope. All these results would lighten the further study of its biological functions in vitro and in vivo.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Recombinant Proteins
/
Immunoglobulin Variable Region
/
Tumor Cells, Cultured
/
Transfection
/
Lymphoma, B-Cell
/
Cricetulus
/
CHO Cells
/
CD3 Complex
/
Antibodies, Bispecific
/
Antigens, CD20
Type of study:
Prognostic study
Limits:
Animals
/
Humans
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2005
Type:
Article
Similar
MEDLINE
...
LILACS
LIS