1. Development of a high-throughput assay to detect antibody inhibition of low pH induced conformational changes of influenza virus hemagglutinin
- Author
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Jacqueline M. Katz, Seiji Hongo, Xiuhua Lu, Zhu-Nan Li, Jessica F. Trost, Paul J. Carney, Kanetsu Sugawara, Terrence M. Tumpey, Elizabeth LeMasters, James Stevens, David A. Steinhauer, Feng Liu, and Min Z. Levine
- Subjects
Models, Molecular ,RNA viruses ,0301 basic medicine ,Conformational change ,Protein Conformation ,Physiology ,Cell Membranes ,lcsh:Medicine ,Hemagglutinin Glycoproteins, Influenza Virus ,medicine.disease_cause ,Biochemistry ,Membrane Fusion ,Physical Chemistry ,law.invention ,Protein structure ,law ,Immune Physiology ,Medicine and Health Sciences ,Influenza A virus ,Enzyme-Linked Immunoassays ,lcsh:Science ,Pathology and laboratory medicine ,Immune System Proteins ,Multidisciplinary ,biology ,Chemistry ,Proteases ,Hydrogen-Ion Concentration ,Medical microbiology ,Recombinant Proteins ,Enzymes ,3. Good health ,Physical Sciences ,Viruses ,Recombinant DNA ,Sorption ,Cellular Structures and Organelles ,Pathogens ,Antibody ,Research Article ,medicine.drug_class ,Immunology ,030106 microbiology ,Hemagglutinin (influenza) ,Enzyme-Linked Immunosorbent Assay ,Research and Analysis Methods ,Monoclonal antibody ,Microbiology ,Antibodies ,Virus ,03 medical and health sciences ,Influenza, Human ,Protein Concentration Assays ,medicine ,Humans ,Influenza viruses ,Immunoassays ,Molecular Biology Techniques ,Molecular Biology ,Molecular Biology Assays and Analysis Techniques ,Influenza A Virus, H3N2 Subtype ,lcsh:R ,Organisms ,Viral pathogens ,Biology and Life Sciences ,Proteins ,Cell Biology ,Antibodies, Neutralizing ,Molecular biology ,High-Throughput Screening Assays ,Microbial pathogens ,030104 developmental biology ,Enzymology ,Immunologic Techniques ,biology.protein ,lcsh:Q ,Adsorption ,Serine Proteases ,Orthomyxoviruses - Abstract
Many broadly neutralizing antibodies (bnAbs) bind to conserved areas of the hemagglutinin (HA) stalk region and can inhibit the low pH induced HA conformational changes necessary for viral membrane fusion activity. We developed and evaluated a high-throughput virus-free and cell-free ELISA based low pH induced HA Conformational Change Inhibition Antibody Detection Assay (HCCIA) and a complementary proteinase susceptibility assay. Human serum samples (n = 150) were tested by HCCIA using H3 recombinant HA. Optical density (OD) ratios of mAb HC31 at pH 4.8 to pH 7.0 ranged from 0.87 to 0.09. Our results demonstrated that low pH induced HA conformational change inhibition antibodies (CCI) neutralized multiple H3 strains after removal of head-binding antibodies. The results suggest that HCCIA can be utilized to detect and characterize CCI in sera, that are potentially broadly neutralizing, and serves as a useful tool for evaluating universal vaccine candidates targeting the HA stalk.
- Published
- 2018