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Updated: Aug 17, 2026
Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood
Published on: February 6, 2018
[Expression of a candidate human Era binding protein A19 and the preparation of its polyclonal antibody]
Yun-feng Li1, Yuan-ming Wu, Su-min Chen
1Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China. Liyf@263.net
Aim:
To express a candidate hEra binding protein A19 in Escherichia coli and to prepare anti-A19 antibody.
Methods:
A19 gene was amplified by PCR from the plasmid containing A19 gene and was cloned into the expression vector pGEX-4T3 which was then transformed into E.coli. The A19 protein was expressed under IPTG induction. Antiserum was prepared by immunizing rabbits with the expressed A19 protein. The titer and specificity of polyclonal antibody were detected by Western blot.
Results:
The expressed A19 accounted for about 30.2% of total bacterial protein. The titer of the antiserum was about 1:4 000. Western blot analysis indicated that the antiserum had high specificity.
Conclusion:
A19 fusion protein was highly expressed. The specific anti-A19 antiserum was prepared successfully.
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