Next Article in Journal
P2X and P2Y Receptors—Role in the Pathophysiology of the Nervous System
Next Article in Special Issue
A Multi-Objective Approach for Protein Structure Prediction Based on an Energy Model and Backbone Angle Preferences
Previous Article in Journal
Emerging Regulation and Function of Betatrophin
Previous Article in Special Issue
Thermal Stability Threshold for Amyloid Formation in Light Chain Amyloidosis
Article Menu
Issue 12 (December) cover image

Export Article

Open AccessArticle
Int. J. Mol. Sci. 2014, 15(12), 23658-23671; doi:10.3390/ijms151223658

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.
Received: 16 September 2014 / Revised: 1 December 2014 / Accepted: 12 December 2014 / Published: 18 December 2014
(This article belongs to the Collection Protein Folding)
View Full-Text   |   Download PDF [1696 KB, uploaded 18 December 2014]   |  

Abstract

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
Figures

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

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.

Show more citation formats Show less citations formats

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top