Development and evaluation of multiplexed immunoassay for detection of antibodies to HPV vaccine types

Details

• Alternative Title:
  J Immunol Methods
• Personal Author:
  Panicker, G. ; Rajbhandari, I. ; Gurbaxani, B.M. ; Querec, T.D. ; Unger, E.R.
• Description:
  Reliable antibody based-assays are needed to evaluate the immunogenicity of current vaccines, impact of altered dosing schemes or of new vaccine formulations. An ideal assay platform would allow multiplex type-specific detection with minimal sample requirement. We used the Meso Scale Discovery (MSD) electrochemiluminescence based detection platform to develop a multiplex direct virus-like particle (VLP) ELISA to detect antibodies to HPV 6, 11, 16, and 18 with a protocol developed for detection using the SI 6000 imager (M4ELISA). MSD prepared the plates in the 7-spot/well format, using the purified VLPs (4 spots) and PBS+BSA pH7.4 (3 blank spots). Three-point titrations and the parallel line method were used to calculate antibody levels. Dynamic range, precision, and stability of pre-printed plates were determined using a panel of previously characterized sera. Cut-off values using children's sera were established using 99% RLU limits based on the 4-parameter Johnson Su best fit curve. Results of the M4ELISA were compared to competitive Luminex Immunoassay (cLIA) on n = 4454 sera from a predominantly unvaccinated cohort. Using a VLP coating concentration of 80 μg/ml with BSA provided the most robust RLU signal for all types. The dynamic range of the assay was about 1000 fold, with assay variability under 25% for each of the four vaccine types. Long-term stability of the plates extended to about 7 months from the time plates was received in the laboratory after printing. There was moderate agreement (κ = 0.38-0.54) between M4ELISA and cLIA, with antibody detection for each of the 4 types more frequent with M4ELISA. Quantitative analysis however showed a good correlation between concordant samples by both assays (ρ ≥ 0.6). The MSD platform shows promise for simultaneous quantitation of the antibody responses to four HPV vaccine types in a high-throughput manner.
• Subjects:
  Antibodies, Viral Article Child Electrochemical Techniques ELISA Enzyme-Linked Immunosorbent Assay Female HPV Antibodies Human Papillomavirus 6 Human Papillomavirus 11 Human Papillomavirus 16 Human Papillomavirus 18 Humans Male Meso Scale Discovery Multiplex Papillomavirus Vaccines Serology

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• Source:
  J Immunol Methods. 417:107-114.
• Pubmed ID:
  25554636
• Pubmed Central ID:
  PMC4587388
• Document Type:
  Journal Article
• Funding:
  CC999999/Intramural CDC HHS/United States
• Volume:
  417
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:140232247ee4313c3e091d9e2174d6e0249a2bff7ba32effdc23839a1a0d503d
• Download URL:
  https://stacks.cdc.gov/view/cdc/34546/cdc_34546_DS1.pdf
• File Type:
  [PDF - 604.34 KB ]