1. Production and characterization of polyclonal and monoclonal antibodies of lamprey pore-forming protein.
- Author
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Nie, Meng, Feng, Bo, Liu, Chang, Tu, Yijun, Chen, Xiaoni, and Wu, Fenfang
- Subjects
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LAMPREYS , *RECOMBINANT proteins , *MONOCLONAL antibodies , *PROTEINS , *HOMODIMERS , *IMMUNOGLOBULINS - Abstract
In the most primitive jawless vertebrate lamprey, the complement-dependent cytotoxicity regulated by variable lymphocyte receptors (VLRs) plays an important role in the adaptive immunity. Our previous studies have shown that the lamprey pore-forming protein (LPFP) acted as the terminal effector of VLR to lyse and kill the target cells. Here, the recombinant GST-LPFP protein was expressed and purified in prokaryotic expression system, and then used as the immunogen to produce mouse monoclonal antibody and rabbit polyclonal antibody. With these antibodies, we proved that LPFP existed as homodimers in the lamprey serum, and could be recruited to the membrane of target cells after stimulation. In conclusion, the antibodies we produced could specifically recognize the LPFP protein, which could be the useful tools to further study the pore-forming mechanism of LPFP. • LPFP protein was successfully expressed and purified via the prokaryotic expression system. • The rabbit polyclonal antibody and mouse monoclonal antibody with very high titers against LPFP were produced. • LPFP existed as a homodimer in the lamprey serum. • LPFP was recruited to the membrane of target cells to directly participate in the complement defense system in lamprey. [ABSTRACT FROM AUTHOR]
- Published
- 2022
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