抗chTLR3单克隆抗体制备及其初步应用

doi:10.11843/j.issn.0366-6964.2019.06.020

Abstract

Abstract: Toll-like receptor 3 (TLR3) is a pattern recognition receptor that stimulates innate immunity. In order to prepare a monoclonal antibody that can bind to chicken TLR3 (chTLR3) protein with natural structure, BALB/c mice were immunized with recombinant plasmid pVAX1-Igk-chTLR3, which could be expressed in cells, and monoclonal antibodies against chTLR3 were prepared by hybridoma technique. The expression of chTLR3 expression of chTLR3 in peripheral blood lymphocytes and spleen of cadmium poisoned chickens were detected by Western blot, flow cytometry and indirect immunofluorescence. The results showed that a positive hybridoma cell line was obtained. The number of chromosomes of the cell line was 94±2, the titer of the cell supernatant was 1:2⁵, and the antibody subclass was identified as IgG3. The cell supernatant not only specifically bound to the prokaryotic expression protein of chTLR3, but also bound to chTLR3 protein in chicken peripheral blood lymphocytes. Western blot, flow cytometry and indirect immunofluorescence were used to detect the expression of chTLR3 in peripheral blood lymphocyte and spleen of chickens in control group and cadmium-exposed group. The results indicated that the monoclonal antibody against chTLR3 with natural activity was successfully prepared. The antibody can be used to study the distribution, localization and expression of chTLR3 in chicken tissues and cells by Western blot, indirect immunofluorescence and flow cytometry. It provides an important material basis for tracing the distribution and expression of chTLR3 protein and its role in the pathogenesis of chicken diseases.

CLC Number:

Q813

TANG Zequn, LUO Haiyan, GE Ming, GUO Rong, XU Danlei, ZHANG Ruili. Preparation of Monoclonal Antibody against chTLR3 and Its Preliminary Application[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(6): 1292-1300.

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Preparation of Monoclonal Antibody against chTLR3 and Its Preliminary Application

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