猪 CD205 分子 CysR-FN Ⅱ蛋白特异性纳米抗体的筛选与鉴定研究.

[Objectives] Porcine CD205, as a receptor specific to porcine dendritic cells (DC), played a key role in the antigen presentation. In this study, nanobodies against porcine CD205 CysR-FN Ⅱ protein were screened by phage display technology, which laid a foundation for the research of porcine CD205-targeted antigen presentation. [Methods] CysR-FN Ⅱ protein of porcine CD205 molecule prepared in the previous research was used for the immunization of alpacas. After subcutaneous immunization for 6 times, peripheral blood lymphocytes of alpaca were collected, their total RNA was extracted and cDNA was obtained by reverse transcription. The gene fragments encoding nanobodies were obtained by PCR amplification, and the porcine CD205 phage display nanobody library was constructed. Bio-spannings were performed by phage display technology and monoclonal colonies were randomly selected to screen specific nanobodies against porcine CD205 CysR-FN Ⅱ protein by Phage-ELISA. [Results] After 4 rounds of bio-spanning, a total of 12 porcine CD205 CysR-FN Ⅱ protein specific nanobodies were selected by Phage-ELISA and sequence analysis with their molecular weight about 17×10³. [Conclusions] This study successfully selected and obtained porcine CD205 CysR-FN Ⅱ protein specific nanobodies. [ABSTRACT FROM AUTHOR]