Prokaryotic expression of the giant panda VEGFA gene and the preparation and characterization of its monoclonal antibodies

Vascular endothelial growth factor A (VEGFA) plays a crucial role in angiogenesis and is closely associated with two major challenges in giant panda (<i>Ailuropoda melanoleuca</i>) conservation: reproductive disorders and cancer. To support the development of diagnostic and therapeutic tools for this vulnerable species, this study aimed to generate and characterize monoclonal antibodies (mAbs) specific to giant panda VEGFA. The VEGFA gene fragment (35-517) was cloned and expressed as a recombinant protein in <i>Escherichia coli</i>, which was used as an immunogen. Through hybridoma technology, four stable mAb-secreting cell lines were established, all producing IgG2b subclass antibodies. The obtained mAbs were first screened and characterized by indirect enzyme-linked immunosorbent assay (ELISA) using the recombinant VEGFA protein. Their specific recognition of the native VEGFA protein in various giant panda tissues and cells was subsequently confirmed by Western blot analysis. In conclusion, this study successfully generated mAbs against giant panda VEGFA, providing a valuable reagent for future studies on pregnancy complications and cancer in this species.