犬白介素31蛋白表达及其单克隆抗体制备OA

Interleukin-31(IL-31)exerts a key biological role in various cutaneous inflammatory processes by binding to its receptor.In this study,the prokaryotic expression vector for canine IL-31 was constructed,and the recombinant canine IL-31 protein with high purity was successfully expressed and purified using an E.coli expression system.After immunizing BALB/c mice with this recombinant protein,followed by three rounds of immunization,cell fusion,and screening via the limited dilution method,one hybridoma cell line 1F2 that could stably secrete canine IL-31 monoclonal antibody was successfully obtained.The antibody was of the IgG1/λ isotype,and the titer of the purified antibody could reach 1∶13 107 200.To further verify the antibody specificity,a eukaryotic expression plasmid was construct-ed and used to express canine IL-31 in HEK-293T cells.Western-blot results showed that the 1F2 mono-clonal antibody could specifically recognize canine IL-31 protein expressed in the eukaryotic system,in-dicating excellent reaction specificity.The successfully prepared anti-canine IL-31 monoclonal antibody 1F2 in this study lays a foundation for the subsequent development of therapeutic antibodies targeting canine IL-31 and the establishment of related detection methods.