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Article

Refolded scFv Antibody Fragment against Myoglobin Shows Rapid Reaction Kinetics

1
Medical Proteomics Research Center, Korea Research Institute of Bioscience & Biotechnology, Daejeon 305-806, Korea
2
Department of Biotechnology and Bioinformatics, Korea University, Sejong 339-700, Korea
3
Department of Food and Biotechnology, Korea University, Sejong 339-700, Korea
4
Department of Bio-analytical Science, University of Science and Technology, Daejeon 305-333, Korea
*
Authors to whom correspondence should be addressed.
Int. J. Mol. Sci. 2014, 15(12), 23658-23671; https://0-doi-org.brum.beds.ac.uk/10.3390/ijms151223658
Received: 16 September 2014 / Revised: 1 December 2014 / Accepted: 12 December 2014 / Published: 18 December 2014
(This article belongs to the Collection Protein Folding)
Myoglobin is one of the early biomarkers for acute myocardial infarction. Recently, we have screened an antibody with unique rapid reaction kinetics toward human myoglobin antigen. Antibodies with rapid reaction kinetics are thought to be an early IgG form produced during early stage of in vivo immunization. We produced a recombinant scFv fragment for the premature antibody from Escherichia coli using refolding technology. The scFv gene was constructed by connection of the VHVL sequence with a (Gly4Ser)3 linker. The scFv fragment without the pelB leader sequence was expressed at a high level, but the solubility was extremely low. A high concentration of 8 M urea was used for denaturation. The dilution refolding process in the presence of arginine and the redox reagents GSH and GSSH successfully produced a soluble scFv protein. The resultant refolded scFv protein showed association and dissociation values of 9.32 × 10−4 M−1·s−1 and 6.29 × 10−3 s−1, respectively, with an affinity value exceeding 107 M−1 (kon/koff), maintaining the original rapid reaction kinetics of the premature antibody. The refolded scFv could provide a platform for protein engineering for the clinical application for diagnosis of heart disease and the development of a continuous biosensor. View Full-Text
Keywords: single-chain variable fragment (scFv); premature antibody; myoglobin; acute myocardial infarction single-chain variable fragment (scFv); premature antibody; myoglobin; acute myocardial infarction
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MDPI and ACS Style

Song, H.-N.; Jang, J.-H.; Kim, Y.-W.; Kim, D.-H.; Park, S.-G.; Lee, M.K.; Paek, S.-H.; Woo, E.-J. Refolded scFv Antibody Fragment against Myoglobin Shows Rapid Reaction Kinetics. Int. J. Mol. Sci. 2014, 15, 23658-23671. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms151223658

AMA Style

Song H-N, Jang J-H, Kim Y-W, Kim D-H, Park S-G, Lee MK, Paek S-H, Woo E-J. Refolded scFv Antibody Fragment against Myoglobin Shows Rapid Reaction Kinetics. International Journal of Molecular Sciences. 2014; 15(12):23658-23671. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms151223658

Chicago/Turabian Style

Song, Hyung-Nam, Jun-Hyuck Jang, Young-Wan Kim, Dong-Hyung Kim, Sung-Goo Park, Myung K. Lee, Se-Hwan Paek, and Eui-Jeon Woo. 2014. "Refolded scFv Antibody Fragment against Myoglobin Shows Rapid Reaction Kinetics" International Journal of Molecular Sciences 15, no. 12: 23658-23671. https://0-doi-org.brum.beds.ac.uk/10.3390/ijms151223658

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