Screening and identification of human anti-C3d single chain antibodies

VernacularTitle: 人源抗C3d单链抗体的筛选与鉴定
Author: Xin-Ping ZHAO ¹
Author Information

1. PLA Center for Disease Control and Prevention
Publication Type:Journal Article
Keywords: Affinity; C3d; Phage display antibody library; Single-chain antibody
From: Journal of International Pharmaceutical Research 2020;47(9):750-755
CountryChina
Language:Chinese
Abstract: Objective: To screen human single chain antibodies against human C3d from phage-displayed single-chain variable fragment(scFv)library, and analyze their binding activities. Methods: The phage display library was exposed to 3-round selections in immunotubes coated with recombinant C3d at a decreasing concentration range and progressively stringent washing conditions. The positive clones were identified by ELISA, followed with sequencing to determine the specific genes which were then cloned intoex-pression vectors. The single chain antibodies were expressed in the FreeStyleTM 293-F system and harvested by affinity purification. The binding activities with recombinant C3d were determined by using Bio-Layer Interferometry technology. Results: These experiments resulted in three novel single chain antibodies(that is, A1, A3 and B6)against C3d with high affinity ranging from 22.7 to 171 pmol/ L. Conclusion: The high affinity human single chain antibodies against human C3d were obtained.