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Development of a high-throughput bead based assay system to measure HIV-1 specific immune signatures in clinical samples


Liechti, Thomas; Kadelka, Claus; Ebner, Hanna; Friedrich, Nikolas; Kouyos, Roger D; Gunthard, Huldrych F; Trkola, Alexandra (2018). Development of a high-throughput bead based assay system to measure HIV-1 specific immune signatures in clinical samples. Journal of Immunological Methods, 454:48-58.

Abstract

The monitoring and assessment of a broadly neutralizing antibody (bnAb) based HIV-1 vaccine require detailed measurements of HIV-1 binding antibody responses to support the detection of correlates of protection. Here we describe the development of a flexible, high-throughput microsphere based multiplex assay system that allows monitoring complex binding antibody signatures. Studying a panel of 13 HIV-1 antigens in a parallel assessment of different IgG subclasses (IgG1, IgG2 and IgG3) we demonstrate the potential of our strategy. The technical advances we describe include means to improve antigen reactivity using directed neutravidin-biotin immobilization of antigens and biotin saturation to reduce background. A particular emphasis of our study was to provide tools for the assessment of reproducibility and stability of the assay system and strategies to control for variations allowing the application in highthroughput assays, where reliability of single measurements needs to be guaranteed.

Abstract

The monitoring and assessment of a broadly neutralizing antibody (bnAb) based HIV-1 vaccine require detailed measurements of HIV-1 binding antibody responses to support the detection of correlates of protection. Here we describe the development of a flexible, high-throughput microsphere based multiplex assay system that allows monitoring complex binding antibody signatures. Studying a panel of 13 HIV-1 antigens in a parallel assessment of different IgG subclasses (IgG1, IgG2 and IgG3) we demonstrate the potential of our strategy. The technical advances we describe include means to improve antigen reactivity using directed neutravidin-biotin immobilization of antigens and biotin saturation to reduce background. A particular emphasis of our study was to provide tools for the assessment of reproducibility and stability of the assay system and strategies to control for variations allowing the application in highthroughput assays, where reliability of single measurements needs to be guaranteed.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > Institute of Medical Virology
04 Faculty of Medicine > University Hospital Zurich > Clinic for Infectious Diseases
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Uncontrolled Keywords:Immunology, Immunology and Allergy
Language:English
Date:2018
Deposited On:19 Jan 2018 12:22
Last Modified:19 Aug 2018 12:58
Publisher:Elsevier
ISSN:0022-1759
Funders:Swiss National Science Foundation (SNF; #314730_152663 and #314730_172790 to AT),, the Clinical Priority Research Program of the University of Zurich (Viral infectious diseases: Zurich Primary HIV Infection Study to HFG and AT),, Swiss Vaccine Research Institute (to AT, HFG, RDK) and the SystemsX.ch grant AntibodyX (to AT)., SNF (#PZ00P3-142411 and BSSGI0_155851) supported RDK, This study has been co-financed within the framework of the Swiss HIV Cohort Study, supported by the SNF (#33CS30_148522 to HFG), by the small nested SHCS project 744 (to AT) and by the SHCS research foundation
OA Status:Green
Publisher DOI:https://doi.org/10.1016/j.jim.2017.12.003
PubMed ID:29277486
Project Information:
  • : FunderSNSF
  • : Grant ID
  • : Project TitleSwiss National Science Foundation (SNF; #314730_152663 and #314730_172790 to AT),
  • : Funder
  • : Grant ID
  • : Project Titlethe Clinical Priority Research Program of the University of Zurich (Viral infectious diseases: Zurich Primary HIV Infection Study to HFG and AT),
  • : Funder
  • : Grant ID
  • : Project TitleSwiss Vaccine Research Institute (to AT, HFG, RDK) and the SystemsX.ch grant AntibodyX (to AT).
  • : FunderSNSF
  • : Grant ID
  • : Project TitleSNF (#PZ00P3-142411 and BSSGI0_155851) supported RDK
  • : FunderSNSF
  • : Grant ID
  • : Project TitleThis study has been co-financed within the framework of the Swiss HIV Cohort Study, supported by the SNF (#33CS30_148522 to HFG), by the small nested SHCS project 744 (to AT) and by the SHCS research foundation

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