Antibody Profiling by Proteome Microarray with Multiplex Isotype Detection Reveals Overlap between Human and Aotus nancymaae Controlled Malaria Infections

Proteomics. 2018 Jan;18(2):10.1002/pmic.201700277. doi: 10.1002/pmic.201700277.

Abstract

The development of vaccines against malaria and serodiagnostic tests for detecting recent exposure requires tools for antigen discovery and suitable animal models. The protein microarray is a high-throughput, sample sparing technique, with applications in infectious disease research, clinical diagnostics, epidemiology, and vaccine development. We recently demonstrated Qdot-based indirect immunofluorescence together with portable optical imager ArrayCAM using single isotype detection could replicate data using the conventional laser confocal scanner system. We developed a multiplexing protocol for simultaneous detection of IgG, IgA, and IgM and compared samples from a controlled human malaria infection model with those from controlled malaria infections of Aotus nancymaae, a widely used non-human primate model of human malaria. IgG profiles showed the highest concordance in number of reactive antigens; thus, of the 139 antigens recognized by human IgG antibody, 111 were also recognized by Aotus monkeys. Interestingly, IgA profiles were largely non-overlapping. Finally, on the path toward wider deployment of the portable platform, we show excellent correlations between array data obtained in five independent laboratories around the United States using the multiplexing protocol (R2 : 0.60-0.92). This study supports the use of this platform for wider deployment, particularly in endemic areas where such a tool will have the greatest impact on global human health.

Keywords: antibody isotype; malaria; multiplex; protein microarray; quantum dots.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Aotidae
  • Fluorescent Antibody Technique, Indirect
  • Humans
  • Immunoassay / methods*
  • Immunoglobulin A / analysis
  • Immunoglobulin G / analysis*
  • Immunoglobulin M / analysis
  • Malaria, Falciparum / diagnosis*
  • Malaria, Falciparum / metabolism
  • Malaria, Falciparum / parasitology
  • Plasmodium falciparum / isolation & purification
  • Protein Array Analysis / methods*
  • Proteome / analysis*
  • Quantum Dots

Substances

  • Immunoglobulin A
  • Immunoglobulin G
  • Immunoglobulin M
  • Proteome