Henry Kevin A, Tanha Jamshid, Hussack Greg
Human Health Therapeutics Portfolio, National Research Council Canada, 100 Sussex Drive, Ottawa, ON, Canada K1A 0R6.
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.
Protein Eng Des Sel. 2015 Oct;28(10):379-83. doi: 10.1093/protein/gzv039. Epub 2015 Aug 27.
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.
与给定蛋白质的多种同工型或同源物发生交叉反应的抗体通常是理想的，特别是在治疗应用中。在此，我们报告了从仅用人血清白蛋白免疫的羊驼中鉴定出几种独特的、克隆无关的单域抗体（sdAb），它们以低至中纳摩尔亲和力结合多种血清白蛋白直系同源物（人、恒河猴、大鼠和小鼠）。通过针对血清白蛋白对噬菌体展示的sdAb文库进行单轮淘选和下一代DNA测序，我们能够以约90%的准确率预测sdAb对不同物种白蛋白的反应模式。我们预计这种策略通常适用于鉴定交叉反应性sdAb，特别是当它们以低频率存在和/或通过淘选难以富集时。此外，这里鉴定出的sdAb对于延长生物制品的血清半衰期具有潜在的意义。
