A cocktail of thermally stable, chemically synthesized capture agents for the efficient detection of anti-gp41 antibodies from human sera

Jessica A Pfeilsticker; Aiko Umeda; Blake Farrow; Connie L Hsueh; Kaycie M Deyle; Jocelyn T Kim; Bert T Lai; James R Heath

doi:10.1371/journal.pone.0076224

A cocktail of thermally stable, chemically synthesized capture agents for the efficient detection of anti-gp41 antibodies from human sera

PLoS One. 2013 Oct 7;8(10):e76224. doi: 10.1371/journal.pone.0076224. eCollection 2013.

Authors

Jessica A Pfeilsticker¹, Aiko Umeda, Blake Farrow, Connie L Hsueh, Kaycie M Deyle, Jocelyn T Kim, Bert T Lai, James R Heath

Affiliation

¹ Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California, United States of America.

Abstract

We report on a method to improve in vitro diagnostic assays that detect immune response, with specific application to HIV-1. The inherent polyclonal diversity of the humoral immune response was addressed by using sequential in situ click chemistry to develop a cocktail of peptide-based capture agents, the components of which were raised against different, representative anti-HIV antibodies that bind to a conserved epitope of the HIV-1 envelope protein gp41. The cocktail was used to detect anti-HIV-1 antibodies from a panel of sera collected from HIV-positive patients, with improved signal-to-noise ratio relative to the gold standard commercial recombinant protein antigen. The capture agents were stable when stored as a powder for two months at temperatures close to 60(o)C.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Antibodies, Anti-Idiotypic / immunology
Click Chemistry / methods
Enzyme-Linked Immunosorbent Assay
Epitopes / immunology
HIV Antibodies / blood
HIV Antibodies / immunology*
HIV Envelope Protein gp41 / immunology*
HIV Infections / blood
HIV Infections / diagnosis
HIV Infections / immunology
HIV Seropositivity / diagnosis
HIV Seropositivity / immunology*
HIV Seropositivity / virology
Humans
Molecular Structure
Peptides / chemical synthesis
Peptides / chemistry
Peptides / immunology*
Protein Binding / immunology
Protein Stability
Signal-To-Noise Ratio
Temperature

Substances

Antibodies, Anti-Idiotypic
Epitopes
HIV Antibodies
HIV Envelope Protein gp41
Peptides
gp41 protein, Human immunodeficiency virus 1

Grants and funding

This work was funded primarily by a grant from the Bill and Melinda Gates Foundation. Additional funding for the development of screening approaches, and for certain capture agent characterization methods, was provided by the Institute for Collaborative Biotechnologies through grant W911NF-09-0001 from the U.S. Army Research Office. The content of the information does not necessarily reflect the position or the policy of the Government, and no official endorsement should be inferred. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.