Preparation and preliminary application of a novel monoclonal antibody against human SIRPα / 第二军医大学学报

ABSTRACT

To prepare a monoclonal antibody against human SIRPα using synthetic peptide, and to use it for immune test to assess its efficacy. Methods: The synthetic peptide of SIRPα was linked with KLH and the product was used as antigen for immunization of BALB/c mice. The mAb anti-SIRPα was obtained by hybridoma technique. The produced mAb was used for flow cytometry, Western blotting analysis and immunohistochemistry assay. Cytokines secreted by PMA-treated THP-1 cells were tested by antibody arrays after exposure to the obtained mAb, and the levels of TNF-α and IL-6 were assayed by ELISA. Results: The mAb secreting hybridoma clone was successfully obtained and it had a satisfactory efficacy when used for flow cytometry, Western blotting analysis and immunohistochemistry assay. Compared with negative control and isotype control, the prepared mAb can stimulate TNF-α and IL-6 secretion in PMA-treated THP-1 cells. Conclusion: We have successfully prepared the mAb against SIRPo using synthetic peptide.