Development and application of a multiplex assay for the simultaneous measurement of antibody responses elicited by common childhood vaccines

Vaccine. 2018 Sep 5;36(37):5600-5608. doi: 10.1016/j.vaccine.2018.07.048. Epub 2018 Aug 4.


Because vaccine co-administration can affect elicited immune responses, it is important to evaluate new vaccines in the context of pre-existing vaccination schedules. This is particularly necessary for new pediatric vaccines, as the World Health Organization's infant immunization program already schedules several vaccines to be administered during the first months of life. To facilitate the assessment of inter-vaccine interference, we developed a pediatric vaccine multiplex assay (PVMA) to simultaneously measure antibodies against vaccines commonly administered to infants, including hepatitis B, Haemophilus influenzae type B, diphtheria, tetanus, pertussis, rubella, and respiratory syncytial virus (RSV). Comparison of antibody concentrations determined by enzyme-linked immunosorbent assays (ELISAs) and the PVMA demonstrated that the PVMA is highly sensitive, specific, reproducible, and accurate. Moreover, the PVMA requires half the time to assess a cohort compared to ELISAs, and only costs marginally more. Demonstrating the utility of the assay, we employed the PVMA to assess vaccine interference in the setting of a candidate vaccine, using the infant HIV vaccines from the completed Pediatric AIDS Clinical Trials Group (PACTG) protocols 230 and 326 as examples. There was no substantial difference in antibody concentrations between vaccine and placebo recipients, which suggests that HIV vaccination did not disrupt antibody responses elicited by routine pediatric vaccines. Thus, the PVMA is a reliable, high-throughput technique that requires minimal sample volume to measure multiple antibody concentrations concurrently, and is an efficient alternative to ELISAs for the measurement of vaccine-elicited antibody responses in large cohorts.

Keywords: Antibody response; HIV vaccines; Multiplex assay; Pediatric vaccines; Vaccine interference.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • AIDS Vaccines / therapeutic use
  • Antibodies, Bacterial / blood*
  • Antibody Formation*
  • Child
  • Child, Preschool
  • Diphtheria-Tetanus-Pertussis Vaccine / therapeutic use
  • Enzyme-Linked Immunosorbent Assay / methods
  • Female
  • Haemophilus Vaccines / therapeutic use
  • Hepatitis B Vaccines / therapeutic use
  • High-Throughput Screening Assays / methods*
  • Humans
  • Immunization Schedule
  • Infant
  • Male
  • Poliovirus Vaccine, Inactivated / therapeutic use
  • Vaccines / therapeutic use*
  • Vaccines, Combined / therapeutic use


  • AIDS Vaccines
  • Antibodies, Bacterial
  • Diphtheria-Tetanus-Pertussis Vaccine
  • Haemophilus Vaccines
  • Hepatitis B Vaccines
  • Poliovirus Vaccine, Inactivated
  • Vaccines
  • Vaccines, Combined