Development of a high-throughput assay to detect antibody inhibition of low pH induced conformational changes of influenza virus hemagglutinin

Details

• Alternative Title:
  PLoS One
• Personal Author:
  Trost, Jessica F. ; LeMasters, Elizabeth H. ; Liu, Feng ; Carney, Paul ; Lu, Xiuhua ; Sugawara, Kanetsu ; Hongo, Seiji ; Stevens, James ; Steinhauer, David A. ; Tumpey, Terrence ; Katz, Jacqueline M. ; Levine, Min Z. ; Li, Zhu-Nan
• Description:
  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.
• Subjects:
  Antibodies, Neutralizing Enzyme-Linked Immunosorbent Assay Hemagglutinin Glycoproteins, Influenza Virus High-Throughput Screening Assays Humans Hydrogen-Ion Concentration Influenza, Human Influenza A Virus, H3N2 Subtype Models, Molecular Protein Conformation Recombinant Proteins

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• Source:
  PLoS One. 13(6).
• Pubmed ID:
  29949635
• Pubmed Central ID:
  PMC6021090
• Document Type:
  Journal Article
• Volume:
  13
• Issue:
  6
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:31f8958d69c78e8d221dad1bbdd53849e9ae048ee0a2d687529be3e39c400654
• Download URL:
  https://stacks.cdc.gov/view/cdc/58842/cdc_58842_DS1.pdf
• File Type:
  [PDF - 5.52 MB ]