A V_L-linker-V_H orientation dependent single chain variable antibody fragment against rabies virus G protein with enhanced neutralizing potency in vivo.

Title:A V_L-linker-V_H orientation dependent single chain variable antibody fragment against rabies virus G protein with enhanced neutralizing potency in vivo.

Volume: 23 Issue: 1

Author(s): Yue Cheng, Zhuang Li, Hualong Xi, Tiejun Gu, Ruosen Yuan, Xiaoxu Chen, Chunlai Jiang, Wei Kong and Yongge Wu

Affiliation:

Keywords: Biotin, HIS tag, rabies virus, truncated RV glycoprotein, V_H and V_L, scFv.

Abstract: Lethal rabies can be prevented effectively by post-exposure prophylactic (PEP) with rabies immunoglobulin (RIG). Single-chain variable fragment (scFv), which is composed of a variable heavy chain (V_H) and variable light chain (V_L) connected by a peptide linker, may be developed as alternative to RIG for neutralizing rabies virus (RV). However, our previously constructed scFv (FV57S) with the (NH2) V_H-linker-V_L (COOH) orientation showed a lower neutralizing potency than its parent RIG. This orientation may inhibit FV57S from refolding into an intact and correct conformation. Therefore, the RFV57S protein with a V_L-linker-V_H orientation was constructed based on FV57S. A HIS tag was incorporated to aid in purification and detection of RFV57S and FV57S. However, abilities of RFV57S and FV57S to bind with the anti-HIS tag mAb were different. Therefore, a novel direct ELISA was established by utilizing a biotin-labeled truncated glycoprotein of RV. Although with similar stability and in vitro neutralizing potency as FV57S, RFV57S showed enhanced binding ability, affinity and in vivo protective efficacy against lethal dose of RV. Our studies support the feasibility of developing a scFv with reversed orientation and provide a novel method for evaluating the binding ability, stability and affinity of engineered antibodies recognizing linear epitope.

Cite this article as:

Cheng Yue, Li Zhuang, Xi Hualong, Gu Tiejun, Yuan Ruosen, Chen Xiaoxu, Jiang Chunlai, Kong Wei and Wu Yongge, A V_L-linker-V_H orientation dependent single chain variable antibody fragment against rabies virus G protein with enhanced neutralizing potency in vivo., Protein & Peptide Letters 2016; 23 (1) . https://dx.doi.org/10.2174/0929866522666151026122752

DOI https://dx.doi.org/10.2174/0929866522666151026122752	Print ISSN 0929-8665
Publisher Name Bentham Science Publisher	Online ISSN 1875-5305