Human-murine chimeric autoantibodies with high affinity and specificity for systemic sclerosis

Frontiers in Immunology

Immunology

Yanhang Zhuo¹  Qin Hong²  Qiong Liang³  Sunhui Chen³ 

[1] Shengli Clinical Medical College of Fujian Medical University, Fujian Provincial Hospital, Fuzhou, China;Center for Experimental Research in Clinical Medicine, Fujian Provincial Hospital, Fuzhou, China;Shengli Clinical Medical College of Fujian Medical University, Fujian Provincial Hospital, Fuzhou, China;Center for Experimental Research in Clinical Medicine, Fujian Provincial Hospital, Fuzhou, China;Fujian Provincial Key Laboratory of Critical Care Medicine, Fujian Provincial Hospital, Fuzhou, China;Shengli Clinical Medical College of Fujian Medical University, Fujian Provincial Hospital, Fuzhou, China;Department of Pharmacy, Fujian Provincial Hospital, Fuzhou, China;

关键词: systemic sclerosis;    ATA;    phage display;    humanized antibody;    antibody modeling;   

DOI  :  10.3389/fimmu.2023.1127849

 received in 2022-12-20, accepted in 2023-06-07,  发布年份 2023

来源: Frontiers

【 摘 要 】

Scleroderma 70 (Scl-70) is commonly used in the clinic for aiding systemic sclerosis (SSc) diagnosis due to its recognition as autoantibodies in the serum of SSc patients. However, obtaining sera positive for anti-Scl-70 antibody can be challenging; therefore, there is an urgent need to develop a specific, sensitive, and easily available reference for SSc diagnosis. In this study, murine-sourced scFv library were screened by phage display technology against human Scl-70, and the scFvs with high affinity were constructed into humanized antibodies for clinical application. Finally, ten high-affinity scFv fragments were obtained. Three fragments (2A, 2AB, and 2HD) were select for humanization. The physicochemical properties of the amino acid sequence, three-dimensional structural basis, and electrostatic potential distribution of the protein surface of different scFv fragments revealed differences in the electrostatic potential of their CDR regions determined their affinity for Scl-70 and expression. Notably, the specificity test showed the half-maximal effective concentration values of the three humanized antibodies were lower than that of positive patient serum. Moreover, these humanized antibodies showed high specificity for Scl-70 in diagnostic immunoassays for ANA. Among these three antibodies, 2A exhibited most positive electrostatic potential on the surface of the CDRs and highest affinity and specificity for Scl-70 but with least expression level; thus, it may provide new foundations for developing enhanced diagnostic strategies for SSc.

【 授权许可】

Unknown   
Copyright © 2023 Chen, Liang, Zhuo and Hong