Preparation and Application of a Monoclonal Antibody Against Chicken TRIM62.

TRIM62 is a member of the tripartite interaction motif (TRIM) family and exerts crucial roles in innate immune response and cancer. To investigate the relationship between its distribution and avian retrovirus replication, in the present study, a monoclonal antibody (mAb) against chicken TRIM62 was generated. The open reading frame of chicken TRIM62 was amplified by polymerase chain reaction (PCR) and inserted into the expression vector pET-28a. The recombinant expression vectors were transformed into Escherichia coli BL21 (DE3). Then recombinant protein His-TRIM62 was expressed under isopropyl-β-D-thiogalactopyranoside (IPTG) induction and purified. BALB/c mice were immunized with the purified recombinant TRIM62 protein. The hybridomas were obtained by fusing mouse myeloma cell line SP2/0 with splenocytes of immunized mice. Western blot showed that the His-TRIM62 and endogenous TRIM62 were recognized by the mAb. The distribution of TRIM62 in no specific pathogen infection (SPF) chickens was detected by immunohistochemistry, and the positive signals of TRIM62 were mainly distributed in vascular endothelial. Our work indicated that the mAb against chicken TRIM62 would be a valuable tool for further study of the role of TRIM62 in avian retrovirus pathogenesis.