Screening and Identification of Antibodies with Cross-binding Activity to Human and Mouse IL-1R3
Objective:Interleukin-1 receptor accessory protein(IL-1R3)is a potential new target for inflammation regulation.The aim of this study is to obtain anti-IL-1R3 antibodies with cross-binding activity to human and mouse IL-1R3 and to lay the foundation for the new drug development and pharmacodynamic mechanism research of a novel inflammation intervention strategy.Methods:Based on the sequence and structure alignment of human and mouse IL-1R3,a phage-displayed human single-chain(scFv)antibody library was challenged by alternately coating immune-tubes with human or mouse IL-1R3.The variable region genes of the resulting antibodies were cloned into eukaryotic expression vectors to prepare antibodies.The binding activities of the candidate antibodies were determined by ELISA and SPR and their functional activities were evaluated by cell assay.A strategy of re-pairing the light and heavy chains of these antibodies was then followed to obtain new antibodies with improved properties.Results:Five antibodies with cross-binding activity to human and mouse IL-1R3 were identified,four of which showed comparable affinity to both human and mouse IL-1R3,approximately 10-7 mol/L.Two candidate antibodies with favorable solubility effectively blocked IL-1R3-mediated activities in the ex vivo functional assay.A new antibody with re-paired light and heavy chains showed improved binding activity and apparently increased solubility.Conclusions:By introducing antibodies AET1907 and 4H6L with comparable human-mouse IL-1R3 cross-binding activity,this study has laid the groundwork for further investigation.
NETL
NSTL
万方数据
