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Production and characterization of single chain Fv directed against beta2-agonist clenbuterol.
J Agric Food Chem. 2006 Sep 06; 54(18):6654-9.JA

Abstract

The single chain Fv (scFv) directed against beta2-agonist clenbuterol (CBL) was produced by using phage display technology. The heavy chain and light chain variable region genes (VH) VL) were amplified by the polymerase chain reaction (PCR) from CBL specific hybridoma cell lines 5D1 and assembled as a single chain Fv (scFv) fragment with linker peptide (Gly4Ser)3. Then the scFv DNA fragment was cloned into M13 phagemid vector pCANTAB5E and the anti-CBL antibody libraries were constructed. Phages displaying scFv were enriched by panning with CBL-ovalbumin (CBL-OVA) conjugate. After only one round of panning, antigen-positive recombinant phage clones were successfully selected by ELISA. The positive phage was used to infect Escherichia coli HB2151, and the expression of soluble scFv was then induced by IPTG. The scFv showed an improved sensitivity (with IC50 of 0.78 +/- 0.005 ng/mL (n = 4)) when compared with the parent monoclonal antibody (MAb) (with IC50 of 1.34 +/- 0.006 ng/mL (n = 4)) in competitive indirect ELISA (CI-ELISA). Cross-reactivity studies showed that the specificity of scFv was similar to that of MAb. The recombinant scFv prepared in this study could be potentially used instead of conventional antisera or MAb for development of a rapid and affordable immunoassay for the detection of residual CBL in biological matrices.

Authors+Show Affiliations

Food Quality and Safety Research Institute, South China Agriculture University, Guangzhou, Guangdong Province, 510642, P. R. China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16939323

Citation

Pan, Ke, et al. "Production and Characterization of Single Chain Fv Directed Against Beta2-agonist Clenbuterol." Journal of Agricultural and Food Chemistry, vol. 54, no. 18, 2006, pp. 6654-9.
Pan K, Wang H, Zhang HB, et al. Production and characterization of single chain Fv directed against beta2-agonist clenbuterol. J Agric Food Chem. 2006;54(18):6654-9.
Pan, K., Wang, H., Zhang, H. B., Liu, H. W., Lei, H. T., Huang, L., & Sun, Y. M. (2006). Production and characterization of single chain Fv directed against beta2-agonist clenbuterol. Journal of Agricultural and Food Chemistry, 54(18), 6654-9.
Pan K, et al. Production and Characterization of Single Chain Fv Directed Against Beta2-agonist Clenbuterol. J Agric Food Chem. 2006 Sep 6;54(18):6654-9. PubMed PMID: 16939323.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Production and characterization of single chain Fv directed against beta2-agonist clenbuterol. AU - Pan,Ke, AU - Wang,Hong, AU - Zhang,Hong-Bin, AU - Liu,Hong-Wei, AU - Lei,Hong-Tao, AU - Huang,Li, AU - Sun,Yuan-Ming, PY - 2006/8/31/pubmed PY - 2006/10/21/medline PY - 2006/8/31/entrez SP - 6654 EP - 9 JF - Journal of agricultural and food chemistry JO - J. Agric. Food Chem. VL - 54 IS - 18 N2 - The single chain Fv (scFv) directed against beta2-agonist clenbuterol (CBL) was produced by using phage display technology. The heavy chain and light chain variable region genes (VH) VL) were amplified by the polymerase chain reaction (PCR) from CBL specific hybridoma cell lines 5D1 and assembled as a single chain Fv (scFv) fragment with linker peptide (Gly4Ser)3. Then the scFv DNA fragment was cloned into M13 phagemid vector pCANTAB5E and the anti-CBL antibody libraries were constructed. Phages displaying scFv were enriched by panning with CBL-ovalbumin (CBL-OVA) conjugate. After only one round of panning, antigen-positive recombinant phage clones were successfully selected by ELISA. The positive phage was used to infect Escherichia coli HB2151, and the expression of soluble scFv was then induced by IPTG. The scFv showed an improved sensitivity (with IC50 of 0.78 +/- 0.005 ng/mL (n = 4)) when compared with the parent monoclonal antibody (MAb) (with IC50 of 1.34 +/- 0.006 ng/mL (n = 4)) in competitive indirect ELISA (CI-ELISA). Cross-reactivity studies showed that the specificity of scFv was similar to that of MAb. The recombinant scFv prepared in this study could be potentially used instead of conventional antisera or MAb for development of a rapid and affordable immunoassay for the detection of residual CBL in biological matrices. SN - 0021-8561 UR - https://www.unboundmedicine.com/medline/citation/16939323/Production_and_characterization_of_single_chain_Fv_directed_against_beta2_agonist_clenbuterol_ L2 - https://dx.doi.org/10.1021/jf060898x DB - PRIME DP - Unbound Medicine ER -