RT Journal Article
SR Electronic
T1 Development of a high-throughput bead based assay system to measure HIV-1 specific immune signatures in clinical samples
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 195362
DO 10.1101/195362
A1 Thomas Liechti
A1 Claus Kadelka
A1 Hanna Ebner
A1 Nikolas Friedrich
A1 Roger D. Kouyos
A1 Huldrych F. Günthard
A1 Alexandra Trkola
YR 2017
UL http://biorxiv.org/content/early/2017/09/28/195362.abstract
AB 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 high-throughput assays, where reliability of single measurements needs to be guaranteed.