Preparation and identification of polyclonal antibodies against Moraxella catarrhalis UspA1.

Hui Wang; Bo Yang; Kesheng Zhao; Jiaji LI; Xin LI; Mengmeng Kong; Chunjie Gong; Yi Wang; Ye Tao; Qiu Zhang; Zheng HU

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Preparation and identification of polyclonal antibodies against Moraxella catarrhalis UspA1.

Author: Hui WANG ¹ ; Bo YANG ¹ ; Kesheng ZHAO ¹ ; Jiaji LI ¹ ; Xin LI ¹ ; Mengmeng KONG ¹ ; Chunjie GONG ¹ ; Yi WANG ¹ ; Ye TAO ¹ ; Qiu ZHANG ¹ ; Zheng HU ¹
Author Information

1. Key Laboratory of Fermentation Engineering, Ministry of Education, School of Food and Biological of Engineering, Hubei University of Technology, Wuhan 430068, Hubei, China.
Publication Type:Journal Article
Keywords: Moraxella catarrhalis; UspA1; polyclonal antibodies; prokaryotic expression
From: Chinese Journal of Biotechnology 2018;34(1):102-109
CountryChina
Language:Chinese
Abstract: To prepare polyclonal antibodies (PcAb) against UspA1 of Moraxella catarrhalis (Mc), we used bioinformatic analysis to determine the surface exposed region in this protein that holds the antigen epitopes. Then the corresponding coding sequences for this fragment was artificially synthesized according to the codon usage of Escherichia coli. The gene fragment was then subcloned into the prokaryotic expression vector pET-28a(+) and expressed in E. coli rosseta (DE3), and then the recombinant UspA1-His proteins were purified. Two New Zealand white rabbits were immunized with this protein to prepare antiserum. The resulting PcAb was then purified from the antiserum with Protein A affinity column. The results of fluorescence antibody assay, enzyme linked immunosorbent assay and Western blotting analysis showed that the PcAb could specifically recognize the surface exposed region of UspA1 on Mc. The preparation of the PcAb laid a foundation of further development of rapid detection technique for M. catarrhalis.