title: Identification of cross-reactive single-domain antibodies against serum albumin using next-generation DNA sequencing.
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.
authors: Human Health Therapeutics Portfolio, National Research Council Canada, 100 Sussex Drive, Ottawa, ON, Canada K1A 0R6.
authors: Kevin A
authors: KA
authors: Henry
authors: Human Health Therapeutics Portfolio, National Research Council Canada, 100 Sussex Drive, Ottawa, ON, Canada K1A 0R6 School of Environmental Sciences, Ontario Agricultural College, University of Guelph, 50 Stone Road East, Guelph, ON, Canada N1G 2W1 Department of Biochemistry, Microbiology and Immunology, University of Ottawa, 75 Laurier Avenue East, Ottawa, ON, Canada K1H 8M5.
authors: Jamshid
authors: J
authors: Tanha
authors: Human Health Therapeutics Portfolio, National Research Council Canada, 100 Sussex Drive, Ottawa, ON, Canada K1A 0R6 greg.hussack@nrc-cnrc.gc.ca.
authors: Greg
authors: G
authors: Hussack
mesh_terms: D000818:Animals; D003429:Cross Reactions; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D051379:Mice; D051381:Rats; D017422:Sequence Analysis, DNA; D012709:Serum Albumin / Q000276:immunology*; D061905:Single-Domain Antibodies / Q000235:genetics* / Q000276:immunology*
chemical_list: D012709:Serum Albumin; D061905:Single-Domain Antibodies
keywords: VHH; next-generation DNA sequencing; phage display; serum albumin; single-domain antibody