Technical Advances of the Recombinant Antibody Microarray Technology Platform for Clinical Immunoproteomics

PLoS One. 2016 Jul 14;11(7):e0159138. doi: 10.1371/journal.pone.0159138. eCollection 2016.

Abstract

In the quest for deciphering disease-associated biomarkers, high-performing tools for multiplexed protein expression profiling of crude clinical samples will be crucial. Affinity proteomics, mainly represented by antibody-based microarrays, have during recent years been established as a proteomic tool providing unique opportunities for parallelized protein expression profiling. But despite the progress, several main technical features and assay procedures remains to be (fully) resolved. Among these issues, the handling of protein microarray data, i.e. the biostatistics parts, is one of the key features to solve. In this study, we have therefore further optimized, validated, and standardized our in-house designed recombinant antibody microarray technology platform. To this end, we addressed the main remaining technical issues (e.g. antibody quality, array production, sample labelling, and selected assay conditions) and most importantly key biostatistics subjects (e.g. array data pre-processing and biomarker panel condensation). This represents one of the first antibody array studies in which these key biostatistics subjects have been studied in detail. Here, we thus present the next generation of the recombinant antibody microarray technology platform designed for clinical immunoproteomics.

MeSH terms

  • Antibodies* / immunology
  • Biomarkers / analysis
  • Biostatistics
  • High-Throughput Screening Assays / methods
  • High-Throughput Screening Assays / standards
  • High-Throughput Screening Assays / statistics & numerical data
  • Humans
  • Immunogenetic Phenomena
  • Protein Array Analysis / methods*
  • Protein Array Analysis / standards
  • Protein Array Analysis / statistics & numerical data
  • Proteomics / methods*
  • Proteomics / standards
  • Proteomics / statistics & numerical data
  • Quality Control
  • Recombinant Proteins / immunology

Substances

  • Antibodies
  • Biomarkers
  • Recombinant Proteins

Grants and funding

This study was supported by research grants from the Swedish Research Council (VR-NT) (CW), VINNOVA (CAKB), and the Foundation of Strategic Research (Strategic Center for Translational Cancer Research—CREATE Health (www.createhealth.lth.se) (CAKB). These funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The funder (Immunovia) provided support in the form of reagents and salaries for authors (LDM, MN, AH), but did not have any additional role in the study design, data collection and analysis, and decision to publish, or preparation of the manuscript.