Glycobiology Research and Training Center, Department of Medicine and Cellular & Molecular Medicine, University of California, San Diego, La Jolla, California, USA.
Nat Biotechnol. 2010 Aug;28(8):863-7. doi: 10.1038/nbt.1651. Epub 2010 Jul 25.
Recombinant glycoprotein therapeutics produced in nonhuman mammalian cell lines and/or with animal serum are often modified with the nonhuman sialic acid N-glycolylneuraminic acid (Neu5Gc; refs. 1,2). This documented contamination has generally been ignored in drug development because healthy individuals were not thought to react to Neu5Gc (ref. 2). However, recent findings indicate that all humans have Neu5Gc-specific antibodies, sometimes at high levels. Working with two monoclonal antibodies in clinical use, we demonstrate the presence of covalently bound Neu5Gc in cetuximab (Erbitux) but not panitumumab (Vectibix). Anti-Neu5Gc antibodies from healthy humans interact with cetuximab in a Neu5Gc-specific manner and generate immune complexes in vitro. Mice with a human-like defect in Neu5Gc synthesis generate antibodies to Neu5Gc after injection with cetuximab, and circulating anti-Neu5Gc antibodies can promote drug clearance. Finally, we show that the Neu5Gc content of cultured human and nonhuman cell lines and their secreted glycoproteins can be reduced by adding a human sialic acid to the culture medium. Our findings may be relevant to improving the half-life, efficacy and immunogenicity of glycoprotein therapeutics.
在非人类哺乳动物细胞系中产生的重组糖蛋白治疗药物，和/或使用动物血清生产的重组糖蛋白治疗药物，通常会被带有非人类唾液酸 N-羟乙酰神经氨酸（Neu5Gc；参考文献 1,2）修饰。在药物开发过程中，这一有文件记录的污染通常被忽视，因为人们认为健康个体不会对 Neu5Gc 产生反应（参考文献 2）。然而，最近的研究结果表明，所有人类都具有 Neu5Gc 特异性抗体，有时抗体水平还很高。通过使用两种临床使用的单克隆抗体进行研究，我们证明了西妥昔单抗（Erbitux）中存在共价结合的 Neu5Gc，但帕尼单抗（Vectibix）中不存在。来自健康人类的抗 Neu5Gc 抗体以 Neu5Gc 特异性方式与西妥昔单抗相互作用，并在体外产生免疫复合物。Neu5Gc 合成具有人类样缺陷的小鼠在注射西妥昔单抗后会产生针对 Neu5Gc 的抗体，并且循环中的抗 Neu5Gc 抗体可以促进药物清除。最后，我们表明，可以通过在培养基中添加人类唾液酸来降低培养的人类和非人类细胞系及其分泌的糖蛋白中的 Neu5Gc 含量。我们的研究结果可能与提高糖蛋白治疗药物的半衰期、疗效和免疫原性有关。
