Built-in RNA-mediated chaperone (chaperna) for antigen folding tailored to immunized hosts.

Abstract

High-quality antibody (Ab) production depends on the availability of immunologically relevant antigens. We present a potentially universal platform for generating soluble antigens from bacterial hosts, tailored to immunized animals for Ab production. A novel RNA-dependent chaperone, in which the target antigen is genetically fused with an RNA-interacting domain (RID) docking tag derived from the immunized host, promotes the solubility and robust folding of the target antigen. We selected the N-terminal tRNA-binding domain of lysyl-tRNA synthetase (LysRS) as the RID for fusion with viral proteins and demonstrated the expression of the RID fusion proteins in their soluble and native conformations; immunization predominantly elicited Ab responses to the target antigen, whereas the "self" RID tag remained nonimmunogenic. Differential immunogenicity of the fusion proteins greatly enriched and simplified the screening of hybridoma clones of monoclonal antibodies (mAbs), enabling specific and sensitive serodiagnosis of MERS-CoV infection. Moreover, mAbs against the consensus influenza hemagglutinin stalk domain enabled a novel assay for trivalent seasonal influenza vaccines. The Fc-mediated effector function was demonstrated, which could be harnessed for the design of next-generation "universal" influenza vaccines. The nonimmunogenic built-in antigen folding module tailored to a repertoire of immunized animal hosts will drive immunochemical diagnostics, therapeutics, and designer vaccines.

• Authors: Young-Seok Kim, Jongkwan Lim, Jemin Sung, Yucheol Cheong, Eun-Young Lee, Jihoon Kim, Hana Oh, Yeon-Sook Kim, Nam-Hyuk Cho, Seongil Choi, Sang-Moo Kang, Jae-Hwan Nam, Wonil Chae, Baik L Seong
• Journal: Biotechnology and bioengineering (Biotechnol Bioeng) Vol. 117 Issue 7 Pg. 1990-2007 (07 2020) ISSN: 1097-0290 [Electronic] United States
• PMID: 32297972 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2020 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals, Inc.

Chemical References

• Antibodies, Monoclonal
• Antibodies, Viral
• Antigens, Viral
• H1N1 virus hemagglutinin
• Hemagglutinin Glycoproteins, Influenza Virus
• Influenza Vaccines
• Molecular Chaperones
• Recombinant Fusion Proteins
• Lysine-tRNA Ligase

Topics

• Animals
• Antibodies, Monoclonal (biosynthesis, immunology, isolation & purification)
• Antibodies, Viral (biosynthesis, immunology, isolation & purification)
• Antigens, Viral (chemistry, genetics, immunology)
• Coronavirus Infections (blood, diagnosis)
• Enzyme-Linked Immunosorbent Assay
• Female
• Hemagglutinin Glycoproteins, Influenza Virus (chemistry, genetics, immunology)
• Humans
• Hybridomas (immunology)
• Immunization
• Influenza Vaccines
• Lysine-tRNA Ligase (chemistry, genetics)
• Mice
• Mice, Inbred BALB C
• Middle East Respiratory Syndrome Coronavirus (immunology)
• Molecular Chaperones
• Protein Conformation
• Protein Domains
• Protein Folding
• Recombinant Fusion Proteins (chemistry, genetics, immunology)
• Serologic Tests
• Solubility