Applications of ribosome display to antibody drug discovery

Abstract

Ribosome display is a polymerase chain reaction-based in vitro display technology that is well suited to the selection and evolution of high affinity antibodies. Both eukaryotic and prokaryotic translation systems have been applied to ribosome display, and the technology's utility has been demonstrated in the antibody isolation process. In particular, ribosome display lends itself to the evolution of functional characteristics, such as potency, of lead candidate antibodies to provide therapeutic antibodies. Large libraries (10(12)) can be rapidly constructed, antibodies selected, and sequence space extensively explored by targeted mutagenesis techniques or by random mutagenesis throughout the antibody sequence. Using such approaches in ribosome display systems lead antibodies derived from phage display or from immunised animals have been improved > 1000-fold in potency within 6 months. This review will discuss the technology and give an insight into how ribosome display is being applied to the antibody lead discovery and optimisation processes.

Selection and affinity maturation of human antibodies against rabies virus from a scFv gene library using ribosome display

“…Secondary phage display library were made from which antibodies with improved affinity were selected. Similarly, mutagenesis libraries of individual antibody clones, isolated by phage display or ribosome display, have been prepared for ribosome display selection, and this can lead to over 1000-fold increases in potency (Osbourn et al, 1996;Baker et al, 2003;Groves and Osbourn, 2005;Hoogenboom, 2002). Compared with phage display and other cell-surface display technologies, ribosome display remains a powerful alternative that has a unique advantage due to its ability to optimize Abs in vitro.…”

Mutational analysis of domain antibodies reveals aggregation hotspots within and near the complementarity determining regions

“…The ability to reliably and systematically select high‐affinity antibodies against diverse antigens has been greatly improved by the development and refinement of several in vitro antibody selection technologies 1‐8. Cell surface display methods (e.g., phage, bacterial, and yeast display) enable rapid screening of large natural and synthetic antibody libraries for both antibody fragments and full‐length antibodies 1‐6.…”

In vitro display technologies reveal novel biopharmaceutics

“…However, high-affinity antibodies can also reduce efficacy due to impaired tissue penetration into solid tumors (66). Affinity increases of 500-to 1000-fold were achieved for ribosome-displayed antibodies with designed and random mutagenesis (67,68). The fully human Ab Belimumab (LymphoStat-B™, chloroamphenicol acetyltransferase (CAT) & Human Genome Sciences) directed against B-lymphocyte stimulator (BLyS, implicated in the pathogenesis of autoimmune diseases) was isolated from a naive human phage display library (69).…”

“…There is a strong trend in emerging medical science to evolve strategies to visualize and control disease-related processes via targeted agents, and in vitro display technologies are utilized to develop targeted therapeutics and diagnostics for various fields such as cancer, autoimmune and inflammatory diseases, and metabolic and allergic disorders (1,2,67,71,72). Using the massive data pool created by genomics and proteomics (73), they also facilitate the discovery of new drug targets and disease-related genes among ϳ20 -25,000 human genes (74).…”