Cell‐free ribosome display and selection of antibodies on arrayed antigens

Abstract

In vitro display technology is a powerful tool for discovery and optimisation of novel antibodies. With increasing demands on various binding molecules in proteomics studies, techniques for a large-scale generation of antibodies or antibody fragments are needed. Here, we describe a novel method for parallel generation of different antibody fragments (scFv) by integrating cell-free ribosome display with array technology. We have demonstrated the procedure by successfully isolating scFv antibodies specific to 16 different cancer biomarkers via a single process. Our results provide proof of principle for multiple production of various scFv antibodies simultaneously.

La présentation sur ribosome

“…Le ribosome display connaît depuis plusieurs années une attention particulière de la communauté scientifique. [11,30]. Les puces protéiques permettront en parallèle le criblage et la sélection de variants vis-à-vis de plusieurs cibles.…”

“…Affilogic, l'une des seules Depuis, le ribosome display a été utilisé fréquemment pour la sélection, l'évolution et même la maturation des anticorps. 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].…”

Generation and Selection of a Panel of Pan-Filovirus Single-Chain Antibodies using Cell-Free Ribosome Display

“…15 It can also be combined with protein microarrays to allow library-versus-library screening for high-throughput generation of antibodies and genomic discovery of protein–protein interactions. 16,17…”

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

“…Ribosome display of antibody libraries has provided a powerful tool for the selection of mAbs and scFvs to important viral pathogens [ 5 – 7 ]. This display technology allows for rapid in vitro selection and evolution of antigen-specific antibodies from large antibody libraries in a cell-free system [ 8 – 11 ] and high affinity antibodies with desired specificity can be isolated by panning on the antigens of interest [ 6 , 12 , 13 ]. Because of small size, homogeneity and ease of genetic manipulation, scFvs offer significant benefits over traditional monoclonal antibodies, such as speed of development, consistency, and ease of optimization [ 14 – 17 ].…”