Identification of cross-reactive single-domain antibodies against serum albumin using next-generation DNA sequencing

Protein Engineering, Design & Selection : PEDS
Kevin A HenryGreg Hussack

Abstract

Antibodies that cross-react with multiple isoforms or homologue of a given protein are often desirable, especially in therapeutic applications. Here, we report the identification of several unique, clonally unrelated, single-domain antibodies (sdAbs) that bind to multiple serum albumin orthologues (human, rhesus, rat and mouse) with low-to-medium nanomolar affinity from a llama immunized only with human serum albumin. Using single-round panning of a phage-displayed sdAb library against serum albumins and next-generation DNA sequencing, we were able to predict patterns of sdAb reactivity to the albumins of different species with ∼90% accuracy. We expect this strategy to be generally applicable for identifying cross-reactive sdAbs, particularly when these exist at low frequency and/or are poorly enriched by panning. Moreover, the sdAbs identified here are of potential interest for serum half-life extension of biologics.

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Citations

Sep 11, 2016·Protein Engineering, Design & Selection : PEDS·Kevin A HenryC Roger MacKenzie
Sep 28, 2016·Protein Engineering, Design & Selection : PEDS·James S Huston
Nov 21, 2018·The Biochemical Journal·Martin A RossottiC Roger MacKenzie
Aug 31, 2019·Molecular Biotechnology·Rose H ReaderKevin C Gough
Feb 9, 2018·Nucleic Acids Research·Arie RyvkinJonathan M Gershoni
Apr 29, 2020·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Henk van FaassenGreg Hussack
Aug 12, 2020·The FEBS Journal·Serge Muyldermans
Dec 7, 2018·BMC Research Notes·Greg HussackKevin A Henry
Mar 23, 2021·The FEBS Journal·Martin A RossottiJamshid Tanha

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