A Cocktail of Thermally Stable, Chemically Synthesized Capture Agents for the Efficient Detection of Anti-Gp41 Antibodies from Human Sera

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 60oC.

A Chemical Epitope‐Targeting Strategy for Protein Capture Agents: The Serine 474 Epitope of the Kinase Akt2

“…Indeed, the PCC agents developed against this epitope exhibit both inhibitory and activating characteristics depending upon the PCC agent structure. Additional advantages relative to the mAb include the well‐defined chemical structure of the PCC agents developed, as well as their stability against proteases and physical stresses 27. Delivery of these or similarly targeted ligands into live cells to explore their potential as therapeutic inhibitors of Akt2 is under investigation.…”

Recent trends in click chemistry as a promising technology for virus-related research

“…It was reported that tetraphenylethylene (TPE) clicked with a fluorescent 6′sialyllactose moiety can be used as a "turn-on" fluorescent sensor to detect the influenza virus (Kato et al, 2010). A cocktail of thermally stable capture agents generated using click chemistry can be used to detect the anti-gp41 antibodies of HIV-1 from human sera with high sensitivity and specificity (Pfeilsticker et al, 2013). To increase the sensitivity of the biosensor, a variable domain of llama heavy-chain antibodies, which can specifically recognize the foot-and-mouth disease virus, was clicked with azides to generate bioorthogonal reactive groups (Trilling et al, 2014).…”

A Chemical Epitope‐Targeting Strategy for Protein Capture Agents: The Serine 474 Epitope of the Kinase Akt2

“…Additional advantages relative to the mAb include the well-defined chemical structure of the PCC agents developed, as well as their stability against proteases and physical stresses. [27] Delivery of these or similarly targeted ligands into live cells to explore their potential as therapeutic inhibitors of Akt2 is under investigation.…”