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Updated: Jun 27, 2026
Single-cell Screening Method for the Selection and Recovery of Antibodies with Desired Specificities from Enriched Human Memory B Cell Populations
Published on: August 22, 2019
[Cloning and prokaryotic expression of recombinant Jo-1 antigen and identification of its antigen specificity]
Xiao-Yu Zhao1, Zhi-Li Bi, Yi-Hong Wu
1College of Life Science, Hebei University, Reseach Center for Bioengineering Technology of Hebei Province, Baoding 071002, China.
Aim:
To obtain highly purified Jo-1 autoantigens.
Methods:
The full length of DNA sequence coding for Jo-1 (histidyl-tRNA synthetase) was obtained from human placenta by RT-PCR and then it was inserted into pTYB11 or pMAL-c to construct the expression vectors pTYB11-Jo-1 and pMAL-c-Jo-1. The recombinant plasmids were transformed into ER2566 and BL21 of E.coli, respectively.
Results:
The fusion Jo-1 antigens were expressed, Western blot analysis demonstrated they responded specifically to anti-Jo-1 antibody from the patients with autoimmune disease polymyositis and dermatomyositis, but did not respond to normal sera and 188 sera containing anti-RNP, Sm, Ro/La or RNP/Ro antibodies from rheumatosis patients.
Conclusion:
The expressed protein of pMAL-c-Jo-1 is soluble, which accounts for more then 50% of total proteins of cells and can be purified by affinity chromatography. The purified proteins can be used as reagents for determining the anti Jo-1 antibody in the serum of patients.
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