High-yield soluble expression of recombinant influenza virus antigens from Escherichia coli and their potential uses in diagnosis.

Abstract

Although antiviral drugs and vaccines have been successful for mitigating influenza virus infections, the lack of general technical platform for the timely supply of soluble and highly purified influenza viral antigens presents a serious bottleneck for the subsequent analysis for the effective control of the viral disease. Using the Escherichia coli (E. coli) lysyl tRNA synthetase (LysRS) as a novel fusion partner, this study reports the soluble expression of influenza viral proteins in E. coli host, construction of antibody library against the virus, and detection of anti-influenza antibodies using the expressed viral antigens. When influenza A and B viral proteins were fused with the LysRS, the fusion proteins were expressed predominantly as soluble forms and their production yields were high enough to be amenable to immunization protocols in rabbits for antibody generation. The produced antibodies showed high level binding specificity against the respective viral proteins, with cross-reactivity across heterologous viruses within the same type of influenza viruses. In addition, LysRS-HA fusion protein could bind specifically to anti-HA antibodies in the virus-infected mouse serum in widely accepted two detection methods, Western blot and ELISA. These results present a convenient tool for the production of antibodies specific to the virus as well as the rapid detection of influenza viral infections, ultimately contributing to the control of influenza viruses including highly pathogenic H5N1, pandemic H1N1, or the recent H7N9 influenza viruses.

• Authors: Yo Han Jang, Seung Hee Cho, Ahyun Son, Yun Ha Lee, Jinhee Lee, Kwang-Hee Lee, Baik Lin Seong
• Journal: Journal of virological methods (J Virol Methods) Vol. 196 Pg. 56-64 (Feb 2014) ISSN: 1879-0984 [Electronic] Netherlands
• PMID: 24216235 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2013 Elsevier B.V. All rights reserved.

Chemical References

• Antibodies, Viral
• Antigens, Viral
• Recombinant Fusion Proteins

Topics

• Animals
• Antibodies, Viral (blood, isolation & purification)
• Antigens, Viral (genetics, immunology, isolation & purification)
• Blotting, Western
• Enzyme-Linked Immunosorbent Assay
• Escherichia coli (genetics)
• Female
• Gene Expression
• Influenza A virus (genetics)
• Influenza B virus (genetics)
• Mice
• Mice, Inbred BALB C
• Rabbits
• Recombinant Fusion Proteins (genetics, immunology, isolation & purification)