Related Experiment Video
Updated: May 4, 2026
Production of Monoclonal Antibodies Targeting Aminopeptidase N in the Porcine Intestinal Mucosal Epithelium
Published on: May 18, 2021
[Expression of chicken pulmonary surfactant protein A and preparation of its polyclonal antibody]
Hongmei Liu1, Hao Wang, Ying Shao
1School of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China.
Objective:
To express chicken pulmonary surfactant protein A (cSP-A) in Escherichia coli and prepare polyclonal antibodies against the recombinant cSP-A.
Methods:
The carbohydrate recognition domain (CRD) gene of cSP-A (cSP-A-CRD) was artificially synthesized, subcloned into the prokaryotic expression vector pGEX-6P-1, and expressed in Escherichia coli BL21(DE3). After the expression products were identified by SDS-PAGE, the gel containing the target protein was cut and further purified. Mice were innoculated with the purified recombinant protein to prepare polyclonal antibodies against cSP-A. The antibody titer was determined by ELISA, and the specificity and application scope of the polyclonal antibodies were analyzed by Western blotting, immunohistochemical fluorescence assay and ELISA.
Results:
The serum against cSP-A was obtained and had an ELISA titer of 1×10(5);. The serum also showed excellent specificity and reactivity as demonstrated by the Western blotting. It could be used to detect the cSP-A in chicken lung epithelial cells by immunohistochemical fluorescence staining and could also be used to detect the hydrosoluble cSP-A in chicken bronchoalveolar lavage fluid by indirect ELISA.
Conclusion:
The recombinant cSP-A was successfully expressed, and its mouse polyclonal antibodies were obtained.
