Drabek Dubravka, Janssens Rick, de Boer Ernie, Rademaker Rik, Kloess Johannes, Skehel John, Grosveld Frank
Department of Cell Biology, Erasmus MC , Rotterdam , Netherlands.
Harbour Antibodies BV , Rotterdam , Netherlands.
Front Immunol. 2016 Dec 19;7:619. doi: 10.3389/fimmu.2016.00619. eCollection 2016.
Several technologies have been developed to isolate human antibodies against different target antigens as a source of potential therapeutics, including hybridoma technology, phage and yeast display systems. For conventional antibodies, this involves either random pairing of VH and variable light (VL) domains in combinatorial display libraries or isolation of cognate pairs of VH and VL domains from human B cells or from transgenic mice carrying human immunoglobulin loci followed by single-cell sorting, single-cell RT-PCR, and bulk cloning of isolated natural VH-VL pairs. Heavy-chain-only antibodies (HCAbs) that naturally occur in camelids require only heavy immunoglobulin chain cloning. Here, we present an automatable novel, high-throughput technology for rapid direct cloning and production of fully human HCAbs from sorted population of transgenic mouse plasma cells carrying a human HCAb locus. Utility of the technique is demonstrated by isolation of diverse sets of sequence unique, soluble, high-affinity influenza A strain X-31 hemagglutinin-specific HCAbs.
已经开发了几种技术来分离针对不同靶抗原的人源抗体，作为潜在治疗药物的来源，包括杂交瘤技术、噬菌体和酵母展示系统。对于传统抗体，这涉及在组合展示文库中随机配对重链可变区（VH）和轻链可变区（VL），或者从人B细胞或携带人免疫球蛋白基因座的转基因小鼠中分离VH和VL结构域的同源对，随后进行单细胞分选、单细胞逆转录聚合酶链反应（RT-PCR）以及对分离的天然VH-VL对进行批量克隆。骆驼科动物中天然存在的仅重链抗体（HCAbs）仅需要重免疫球蛋白链克隆。在这里，我们展示了一种可自动化的新型高通量技术，用于从携带人HCAb基因座的转基因小鼠浆细胞分选群体中快速直接克隆和生产完全人源的HCAbs。通过分离多种独特序列、可溶、高亲和力的甲型流感病毒X-31株血凝素特异性HCAbs，证明了该技术的实用性。
