Evaluation of the cyclic single-chain Fv antibody derived from nivolumab by biophysical analyses and in vitro cell-based bioassay

Author:

Kamesawa Sena1,Ogawa Mizuki1,Funakoshi Yoshiki1,Kato Masaya1,Kai Shosei1,Namikawa Mana1,Okazaki Kyo1,Sato Takashi1,Kobashigawa Yoshihiro1,Morioka Hiroshi1

Affiliation:

1. Kumamoto University Department of Analytical and Biophysical Chemistry, Graduate School of Pharmaceutical Sciences, , 5-1 Oe-honmachi, Chuo-ku, Kumamoto 862-0973, Japan

Abstract

Abstract Single-chain Fv (scFv) is a recombinant small antibody in which a polypeptide linker connects the variable regions of the light chain (VL) and the heavy chain (VH). The practical use of scFv, however, has been prevented by its tendency to aggregate due to interchain VL–VH interactions. We recently developed a cyclic scFv whose N-terminus and C-terminus were connected by protein ligation techniques. Biophysical comparisons between cyclic and linear scFv have been conducted, but cell biological evaluations remain unexplored. Here we studied the properties of cyclic and linear scFv derived from nivolumab. Biophysical studies revealed that the thermal stability was not changed but that the antigen-binding activity was approximately 3-fold higher as a result of circularization. A cell-based PD-1/PD-L1 interaction inhibitory assay revealed that the biological activity of scFv was markedly higher in the circularized form. In addition, biophysical analysis of scFv proteins incubated in the presence of serum revealed that circularization suppressed the decrease in antigen-binding activity. It could be assumed that circularization of scFv improved stability in the presence of serum, which in turn would suggest the applicability of cyclic scFv as a biopharmaceutical format.

Funder

Japan Agency for Medical Research and Development

Publisher

Oxford University Press (OUP)

Reference30 articles.

1. Bispecific antibodies rise again;Garber;Nat. Rev. Drug Discov.,2014

2. Bispecific antibodies;Kontermann;Drug Discov. Today,2015

3. Bispecific antibodies for cancer therapy: a review;Krishnamurthy;Pharmacol. Ther.,2018

4. Bispecific antibodies: design, therapy, perspectives;Sedykh;Drug Des. Devel. Ther.,2018

5. Antibody–drug conjugates;Zolot;Nat. Rev. Drug Discov.,2013

同舟云学术

1.学者识别学者识别

2.学术分析学术分析

3.人才评估人才评估

"同舟云学术"是以全球学者为主线,采集、加工和组织学术论文而形成的新型学术文献查询和分析系统,可以对全球学者进行文献检索和人才价值评估。用户可以通过关注某些学科领域的顶尖人物而持续追踪该领域的学科进展和研究前沿。经过近期的数据扩容,当前同舟云学术共收录了国内外主流学术期刊6万余种,收集的期刊论文及会议论文总量共计约1.5亿篇,并以每天添加12000余篇中外论文的速度递增。我们也可以为用户提供个性化、定制化的学者数据。欢迎来电咨询!咨询电话:010-8811{复制后删除}0370

www.globalauthorid.com

TOP

Copyright © 2019-2024 北京同舟云网络信息技术有限公司
京公网安备11010802033243号  京ICP备18003416号-3