Ribosome display for the rapid generation of high-affinity Zika-neutralizing single-chain antibodies

Abstract

BackgroundZika virus (ZIKV) is an emerging pathogen with no approved therapeutics and only limited diagnostics available. To address this gap, six mouse single-chain antibodies (scFvs) to ZIKV envelope (E) protein were isolated rapidly and efficiently from a ribosome-displayed antibody library constructed from the spleens of five immunized mice.Methodology/ResultsIn this report, we have generated a panel of mouse scFvs to ZIKV E protein using ribosome display. The six scFvs demonstrated no cross-reactivity with DENV2 NGC envelope protein, suggesting specificity for ZIKV E protein. These scFvs showed differences in their affinity: two (scFv45-3, scFv63-1) of them were dominant after four rounds of panning, and showed higher affinity (an apparent Kd values from 19 to 27 nM) than the other four (scFv5-1, scFv7-2, scFv38-1, and scFv51-2). All six scFvs showed ZIKV-neutralizing activity in the plaque reduction neutralization test (PRNT) assay and their neutralizing activity was positively correlated with their affinities.Conclusions/SignificanceThe scFvs (45–3 and 63–1) with highest affinity may have dual utility as diagnostics capable of recognizing ZIKV E subtypes and may be further developed to treat ZIKV infection. Our approach has the added advantage of generating Fc receptor-deficient antibodies, minimizing concern of antibody-dependent enhancement (ADE) of infection.

Ribosome Display Technology: Applications in Disease Diagnosis and Control

“…There are no approved vaccines yet for this disease and diagnostics were, until recently, quite limited. To bridge these gaps, our laboratory utilized ribosome display to generate high-affinity scFv antibodies that have specificity to ZIKV envelope proteins [ 101 ]. In addition to their high-affinity binding and specificity, these scFvs were also able to neutralize live ZIKV and inhibit infectivity [ 101 ].…”

“…To bridge these gaps, our laboratory utilized ribosome display to generate high-affinity scFv antibodies that have specificity to ZIKV envelope proteins [ 101 ]. In addition to their high-affinity binding and specificity, these scFvs were also able to neutralize live ZIKV and inhibit infectivity [ 101 ]. These single-chain antibodies have great potential and could serve as diagnostics or treatments of ZIKV infections ( Figure 3 ).…”

“… Zika envelope binding and neutralizing analysis of scFvs (adapted from Kunamneni et al [ 101 ]). ( a ) Binding curve of scFvs to Zika E by ELISA.…”

Antibodies: Will Their Star Continue to Rise?

“…A recent Phase 1b clinical trial has suggested that a combination of ‘broadly neutralizing antibodies’ is an improvement on current HIV therapies [4], and six Zika antibodies have been produced for utility in testing for and treating the virus [5]. The Pandemic Prevention Program, a co-operation between DARPA and a number of US-based institutions, is seeking to use synthetic biology to turn patients themselves into ‘antibody factories’.…”

La présentation sur ribosome

“…Une étude de 2016 a par exemple révélé la puissance du ribosome display couplé aux puces protéiques pour cribler à moyen débit plusieurs fragments d'anticorps contre une vingtaine d'antigènes différents[11]. Plus récemment, une étude a permis la génération d'un anticorps neutralisant contre le virus Zika[12].…”