Parameters and determinants of responses to selection in antibody libraries

Author summary Antibodies in the immune system consist of a genetically encoded scaffold that exposes a few highly diverse, non-genetically encoded sites. This focused diversity is sufficient to produce antibodies that bind to any target molecule. To understand the role of the scaffold, which acquires hypermutations during the immune response, over the selective response, we analyze quantitative in vitro experiments where large antibody populations based on different scaffolds are selected against different targets. We show that selective responses are described statistically by two parameters, one of which depends on prior evolution of the scaffold as part of a previous response. Our work provides methods to assay whether naive antibody scaffolds are endowed with a distinctively high selective potential. The sequences of antibodies from a given repertoire are highly diverse at few sites located on the surface of a genome-encoded larger scaffold. The scaffold is often considered to play a lesser role than highly diverse, non-genome-encoded sites in controlling binding affinity and specificity. To gauge the impact of the scaffold, we carried out quantitative phage display experiments where we compare the response to selection for binding to four different targets of three different antibody libraries based on distinct scaffolds but harboring the same diversity at randomized sites. We first show that the response to selection of an antibody library may be captured by two measurable parameters. Second, we provide evidence that one of these parameters is determined by the degree of affinity maturation of the scaffold, affinity maturation being the process by which antibodies accumulate somatic mutations to evolve towards higher affinities during the natural immune response. In all cases, we find that libraries of antibodies built around maturated scaffolds have a lower response to selection to other arbitrary targets than libraries built around germline-based scaffolds. We thus propose that germline-encoded scaffolds have a higher selective potential than maturated ones as a consequence of a selection for this potential over the long-term evolution of germline antibody genes. Our results are a first step towards quantifying the evolutionary potential of biomolecules.