| Status | 已發表Published |
| Soluble expression, purification and bioactivity assay of anti-ovarian carcinoma x anti-CD3 x anti-CD28VH single chain trispecific antibody | |
Zhao, B.; Wang, X.; Zhao, Q.; Wang, S.; Huang, H.
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| 2004-06-01 | |
| Source Publication | Chinese Journal of Microbiology and Immunology
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| ISSN | 0254-5101 |
| Pages | 475-479 |
| Abstract | Objective: To investigate the soluble expression and to purify a recombinant multifunctional single chain trispecific antibody (scTsAb) which contains anti-ovarian carcinoma(oc) scFv, anti-CD3 scFv and VH domain of anti-CD28 antibody and in vitro bioactivity of the purified scTsAb. Methods: The plasmid of scTsAb was transformed into E. coli strain BL21(DE3) Star. The scTsAb was over-expressed as a soluble protein when the E. coli was induced in a low concentration of IPTG (0.2mmol/L) and cultured at a low temperature (30°C). The scTsAb was purified by DEAE anion exchange chromatography. ELISA and FACS mensurate in vitro bioactivity of the purified scTsAb. Results: (1) The soluble expression ratio is about 56%. (2) The scTsAb flow through DEAE chromatography and the purity is over 90%. (3) The purified scTsAb shows strong binding activities to the antigen of ovarian carcinoma cells, recombinant CD28 pure antigen and to CD3 molecules on Jurkat cells. (4) The purified antibody tightly binds to Jurkat cells and SKOV3 cells. Conclusion: The soluble scTsAb maintaines its inherent bioactivity after purification by DEAE anion exchange chromatography. These results provide a technical guarantee to produce abundant sample for further bioactivity experiments and in vivo experiments. They also provide the reference standard for expression, purification and production of similar genetical engineered antibodies. |
| Keyword | Bispecific Antibodies Blinatumomab Single Chain Fragment Variable Antibody |
| URL | View the original |
| Language | 英語English |
| The Source to Article | PB_Publication |
| PUB ID | 55349 |
| Fulltext Access | |
| Document Type | Journal article |
| Collection | DEPARTMENT OF BIOMEDICAL SCIENCES |
| Corresponding Author | Huang, H. |
| Recommended Citation GB/T 7714 | Zhao, B.,Wang, X.,Zhao, Q.,et al. Soluble expression, purification and bioactivity assay of anti-ovarian carcinoma x anti-CD3 x anti-CD28VH single chain trispecific antibody[J]. Chinese Journal of Microbiology and Immunology, 2004, 475-479. |
| APA | Zhao, B.., Wang, X.., Zhao, Q.., Wang, S.., & Huang, H. (2004). Soluble expression, purification and bioactivity assay of anti-ovarian carcinoma x anti-CD3 x anti-CD28VH single chain trispecific antibody. Chinese Journal of Microbiology and Immunology, 475-479. |
| MLA | Zhao, B.,et al."Soluble expression, purification and bioactivity assay of anti-ovarian carcinoma x anti-CD3 x anti-CD28VH single chain trispecific antibody".Chinese Journal of Microbiology and Immunology (2004):475-479. |
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