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[Construction and expression of single-chain antibody of neutralizing monoclonal antibody 13D8 against hepatitis E virus].
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2004 Sep; 20(5):556-9.XB

Abstract

AIM

To weaken the immunogenicity of the neutralizing monoclonal antibody (mAb) 13D8 against hepatitis E virus and express its scFv.

METHODS

The V(L) and V(H) genes were cloned by RT-PCR from hybridoma cells producing mouse mAb. And then V(H)-linker-V(L) fragment (scFv) was constructed and cloned into vector pTO-T7. The scFv protein was expressed in E.coli. The activity of expressed scFv was detected by ELISA and Western blot.

RESULTS

SDS-PAGE analysis showed that the scFv was highly expressed mostly in the form of inclusion body in E.coli, and the yield was up to 26.8% of the total bacteria protein. The results of indirect ELISA and Western blot showed that the expressed scFv could bind specifically to a recombinant protein in OFR2 region of HEV (NE2). The result of competitive ELISA demonstrated that the epitope recognized by the scFv was the same as that by mAb 13D8.

CONCLUSION

The scFv constructed from V(H) and V(L) genes of mAb 13D8 with immunological activity was successfully expressed.

Authors+Show Affiliations

The Key Laboratory of the Ministry of Education for Cell Biology and Tumor Cell Engineering, Xiamen University, Xiamen 361005, China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo 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

15367346

Citation

Li, Li-feng, et al. "[Construction and Expression of Single-chain Antibody of Neutralizing Monoclonal Antibody 13D8 Against Hepatitis E Virus]." Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi = Chinese Journal of Cellular and Molecular Immunology, vol. 20, no. 5, 2004, pp. 556-9.
Li LF, Luo WX, Gu Y, et al. [Construction and expression of single-chain antibody of neutralizing monoclonal antibody 13D8 against hepatitis E virus]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2004;20(5):556-9.
Li, L. F., Luo, W. X., Gu, Y., Zhu, Z. H., Wang, Y. B., Zhang, J., & Xia, N. S. (2004). [Construction and expression of single-chain antibody of neutralizing monoclonal antibody 13D8 against hepatitis E virus]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi = Chinese Journal of Cellular and Molecular Immunology, 20(5), 556-9.
Li LF, et al. [Construction and Expression of Single-chain Antibody of Neutralizing Monoclonal Antibody 13D8 Against Hepatitis E Virus]. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2004;20(5):556-9. PubMed PMID: 15367346.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Construction and expression of single-chain antibody of neutralizing monoclonal antibody 13D8 against hepatitis E virus]. AU - Li,Li-feng, AU - Luo,Wen-xin, AU - Gu,Ying, AU - Zhu,Zi-heng, AU - Wang,Ying-bin, AU - Zhang,Jun, AU - Xia,Ning-shao, PY - 2004/9/16/pubmed PY - 2006/4/6/medline PY - 2004/9/16/entrez SP - 556 EP - 9 JF - Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology JO - Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi VL - 20 IS - 5 N2 - AIM: To weaken the immunogenicity of the neutralizing monoclonal antibody (mAb) 13D8 against hepatitis E virus and express its scFv. METHODS: The V(L) and V(H) genes were cloned by RT-PCR from hybridoma cells producing mouse mAb. And then V(H)-linker-V(L) fragment (scFv) was constructed and cloned into vector pTO-T7. The scFv protein was expressed in E.coli. The activity of expressed scFv was detected by ELISA and Western blot. RESULTS: SDS-PAGE analysis showed that the scFv was highly expressed mostly in the form of inclusion body in E.coli, and the yield was up to 26.8% of the total bacteria protein. The results of indirect ELISA and Western blot showed that the expressed scFv could bind specifically to a recombinant protein in OFR2 region of HEV (NE2). The result of competitive ELISA demonstrated that the epitope recognized by the scFv was the same as that by mAb 13D8. CONCLUSION: The scFv constructed from V(H) and V(L) genes of mAb 13D8 with immunological activity was successfully expressed. SN - 1007-8738 UR - https://www.unboundmedicine.com/medline/citation/15367346/[Construction_and_expression_of_single_chain_antibody_of_neutralizing_monoclonal_antibody_13D8_against_hepatitis_E_virus]_ DB - PRIME DP - Unbound Medicine ER -