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[Construction and expression of single chain variable fragments (ScFv) against human CD19 antigen].
Sheng Wu Gong Cheng Xue Bao. 2005 Sep; 21(5):686-91.SW

Abstract

The genes encoding for the light and heavy chain variable regions were cloned by RT-PCR from a murine monoclonal hybridoma cell line, which could produce monoclonal antibody to recognize CD19 antigen on human B lymphocyte. Then fused the light and heavy chain variable regions together by a short peptide linker containing 15 amino acid (Gly4Ser)3 using splice-overlap extensive PCR. The recombinant anti-CD19- ScFv was subcloned into the expression vector pET28a and induced to be expressed by IPTG in E. coli BL21. SDS-PAGE and Western blot analysis showed that the recombinant anti-CD19-ScFv gene was expressed in E. coli BL21. ScFv expression was in the form of an inclusion bodies and the purified fusion protein was obtained after a series of purification steps including cell break, inclusion body solubilization, Ni2+ metal affinity chromatography and protein refolding. Flow cytometry analysis showed that the ScFv can react with human CD19 antigen. In conclusion, recombinant anti-CD19-ScFv gene has been successful constructed and expressed in E. coli BL21, which could provide a basic study for the future target therapy to the B lymphoid leukemia and B lymphoma.

Authors+Show Affiliations

Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Tianjin 300020, China.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

English Abstract
Journal Article
Research Support, Non-U.S. Gov't

Language

chi

PubMed ID

16285505

Citation

Chen, Sen, et al. "[Construction and Expression of Single Chain Variable Fragments (ScFv) Against Human CD19 Antigen]." Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology, vol. 21, no. 5, 2005, pp. 686-91.
Chen S, Rao Q, Wang JX, et al. [Construction and expression of single chain variable fragments (ScFv) against human CD19 antigen]. Sheng Wu Gong Cheng Xue Bao. 2005;21(5):686-91.
Chen, S., Rao, Q., Wang, J. X., & Wang, M. (2005). [Construction and expression of single chain variable fragments (ScFv) against human CD19 antigen]. Sheng Wu Gong Cheng Xue Bao = Chinese Journal of Biotechnology, 21(5), 686-91.
Chen S, et al. [Construction and Expression of Single Chain Variable Fragments (ScFv) Against Human CD19 Antigen]. Sheng Wu Gong Cheng Xue Bao. 2005;21(5):686-91. PubMed PMID: 16285505.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Construction and expression of single chain variable fragments (ScFv) against human CD19 antigen]. AU - Chen,Sen, AU - Rao,Qing, AU - Wang,Jian-Xiang, AU - Wang,Min, PY - 2005/11/16/pubmed PY - 2009/7/25/medline PY - 2005/11/16/entrez SP - 686 EP - 91 JF - Sheng wu gong cheng xue bao = Chinese journal of biotechnology JO - Sheng Wu Gong Cheng Xue Bao VL - 21 IS - 5 N2 - The genes encoding for the light and heavy chain variable regions were cloned by RT-PCR from a murine monoclonal hybridoma cell line, which could produce monoclonal antibody to recognize CD19 antigen on human B lymphocyte. Then fused the light and heavy chain variable regions together by a short peptide linker containing 15 amino acid (Gly4Ser)3 using splice-overlap extensive PCR. The recombinant anti-CD19- ScFv was subcloned into the expression vector pET28a and induced to be expressed by IPTG in E. coli BL21. SDS-PAGE and Western blot analysis showed that the recombinant anti-CD19-ScFv gene was expressed in E. coli BL21. ScFv expression was in the form of an inclusion bodies and the purified fusion protein was obtained after a series of purification steps including cell break, inclusion body solubilization, Ni2+ metal affinity chromatography and protein refolding. Flow cytometry analysis showed that the ScFv can react with human CD19 antigen. In conclusion, recombinant anti-CD19-ScFv gene has been successful constructed and expressed in E. coli BL21, which could provide a basic study for the future target therapy to the B lymphoid leukemia and B lymphoma. SN - 1000-3061 UR - https://www.unboundmedicine.com/medline/citation/16285505/[Construction_and_expression_of_single_chain_variable_fragments__ScFv__against_human_CD19_antigen]_ DB - PRIME DP - Unbound Medicine ER -