Next Article in Journal
Synthesis of 11C-Labelled Ureas by Palladium(II)-Mediated Oxidative Carbonylation
Previous Article in Journal
PrLPAAT4, a Putative Lysophosphatidic Acid Acyltransferase from Paeonia rockii, Plays an Important Role in Seed Fatty Acid Biosynthesis
Previous Article in Special Issue
NMR Detection of Semi-Specific Antibody Interactions in Serum Environments
Article Menu
Issue 10 (October) cover image

Export Article

Open AccessFeature PaperArticle
Molecules 2017, 22(10), 1695; doi:10.3390/molecules22101695

Production of Single-Chain Fv Antibodies Specific for GA-Pyridine, an Advanced Glycation End-Product (AGE), with Reduced Inter-Domain Motion

1
Department of Analytical and Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Chuo-ku, Kumamoto 862-0973, Japan
2
Department of Molecular Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, 2-2-1 Honjo, Chuo-ku, Kumamoto 860-0811, Japan
3
CREST, JST, 4-1-8, Honcho, Kawaguchi, Saitama 332-0012, Japan
4
Graduate School of Environmental Earth Science, Hokkaido University, Kita-10 Nishi-5, Kita-ku, Sapporo 060-0810, Japan
5
Department of Cell Pathology, Graduate School of Medical Sciences, Kumamoto University, 1-1-1 Honjo, Chuo-ku, Kumamoto 860-8556, Japan
6
Graduate School of Pharmaceutical Sciences, Hokkaido University, Kita-12 Nishi-6, Kita-ku, Sapporo 060-0812, Japan
7
Global Station of Soft Matter, Global Institution for Collaborative Research and Education, Hokkaido University, Kita-15 Nishi-8, Kita-ku, Sapporo 060-0815, Japan
8
Laboratory of Molecular Biophysics, Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan
9
Division of Materials Science and Chemical Engineering, Graduate School of Engineering, Yokohama National University, 79-5 Tokiwadai, Hodogaya-ku, Yokohama 240-8501, Japan
Present address: Kentaro Noi, Department of Mechanical Science and Bioengineering, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama, Toyonaka 560-8531, Japan
Present address: Naoki Ichikawa-Tomikawa, Department of Basic Pathology, Fukushima Medical University School of Medicine, 1 Hikariga-oka, Fukushima 960-1295, Japan
§
Present address: Motoyoshi Nomizu, Department of Clinical Biochemistry, Tokyo University of Pharmacy and Life Sciences, 1432-1 Horinouchi, Hachioji, Tokyo 192-0392, Japan
Present address: Takashi Saito, Division of Pharmaceutics, Hokkaido Pharmaceutical University School of Pharmacy, 7-15-4-1 Maeda, Teine, Sapporo 006-8590, Japan
*
Author to whom correspondence should be addressed.
Received: 15 September 2017 / Revised: 8 October 2017 / Accepted: 9 October 2017 / Published: 10 October 2017
(This article belongs to the Special Issue Recent Advances in Biomolecular NMR Spectroscopy)
View Full-Text   |   Download PDF [3860 KB, uploaded 11 October 2017]   |  

Abstract

Due to their lower production cost compared with monoclonal antibodies, single-chain variable fragments (scFvs) have potential for use in several applications, such as for diagnosis and treatment of a range of diseases, and as sensor elements. However, the usefulness of scFvs is limited by inhomogeneity through the formation of dimers, trimers, and larger oligomers. The scFv protein is assumed to be in equilibrium between the closed and open states formed by assembly or disassembly of VH and VL domains. Therefore, the production of an scFv with equilibrium biased to the closed state would be critical to overcome the problem in inhomogeneity of scFv for industrial or therapeutic applications. In this study, we obtained scFv clones stable against GA-pyridine, an advanced glycation end-product (AGE), by using a combination of a phage display system and random mutagenesis. Executing the bio-panning at 37 °C markedly improved the stability of scFvs. We further evaluated the radius of gyration by small-angle X-ray scattering (SAXS), obtained compact clones, and also visualized open View Full-Text
Keywords: GA-pyridine; single-chain variable fragment; phage display; isothermal titration calorimetry; differential scanning calorimetry; small-angle X-ray scattering; high-speed atomic force microscopy; NMR analysis; inter-domain motion GA-pyridine; single-chain variable fragment; phage display; isothermal titration calorimetry; differential scanning calorimetry; small-angle X-ray scattering; high-speed atomic force microscopy; NMR analysis; inter-domain motion
Figures

Figure 1

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).

Supplementary material

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

Fukuda, N.; Noi, K.; Weng, L.; Kobashigawa, Y.; Miyazaki, H.; Wakeyama, Y.; Takaki, M.; Nakahara, Y.; Tatsuno, Y.; Uchida-Kamekura, M.; Suwa, Y.; Sato, T.; Ichikawa-Tomikawa, N.; Nomizu, M.; Fujiwara, Y.; Ohsaka, F.; Saitoh, T.; Maenaka, K.; Kumeta, H.; Shinya, S.; Kojima, C.; Ogura, T.; Morioka, H. Production of Single-Chain Fv Antibodies Specific for GA-Pyridine, an Advanced Glycation End-Product (AGE), with Reduced Inter-Domain Motion. Molecules 2017, 22, 1695.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]

Molecules EISSN 1420-3049 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top