含有非人唾液酸Neu5Gc残基的重组糖蛋白免疫原性评估的鉴别方法建立。
Method establishment for discerned immunogenicity assessment of a recombinant glycoprotein containing nonhuman sialic acid Neu5Gc residues.
作者信息
Montjovent Marc-Olivier, Zou Linglong, Faust Heinrich, Abdolzade-Bavil Afsaneh, Liu Patrick M, Struwe Petra
机构信息
Celerion Switzerland AG, 8320 Fehraltorf, Switzerland.
Biologics, Assays & Technology, Teva Pharmaceuticals, Inc., West Chester, PA 19380, USA.
出版信息
Bioanalysis. 2017 Sep;9(18):1385-1393. doi: 10.4155/bio-2017-0088.
AIM
Recombinant glycoprotein produced in nonhuman mammalian cell lines can be modified with the immunogenic nonhuman sialic N-glycolylneuraminic acid (Neu5Gc). We describe here a validated method for detection of antidrug antibodies against both protein and Neu5Gc-containing glycan epitopes.
RESULTS
An electrochemiluminescent method was established with drug conjugates as capture and detection reagents. Rabbit antidrug polyclonal antibodies were used as the positive control for protein moiety-specific antibodies, while chicken anti-Neu5Gc polyclonal antibodies were used as the positive control for antibodies against Neu5Gc glycan epitope. Specificity to Neu5Gc was verified by signal inhibition with bovine γ-globulin that contains Neu5Gc.
CONCLUSION
The assay illustrated here discerns the immunogenicity of the protein backbone and the sialic acid Neu5Gc glycan moiety of a recombinant protein containing Neu5Gc.
目的
在非人类哺乳动物细胞系中产生的重组糖蛋白可能会被具有免疫原性的非人类唾液酸N-羟乙酰神经氨酸（Neu5Gc）修饰。我们在此描述一种经过验证的方法，用于检测针对蛋白质和含Neu5Gc聚糖表位的抗药物抗体。
结果
建立了一种以药物偶联物作为捕获和检测试剂的电化学发光方法。兔抗药物多克隆抗体用作蛋白质部分特异性抗体的阳性对照，而鸡抗Neu5Gc多克隆抗体用作抗Neu5Gc聚糖表位抗体的阳性对照。通过用含有Neu5Gc的牛γ球蛋白进行信号抑制来验证对Neu5Gc的特异性。
结论
此处阐述的检测方法能够区分含有Neu5Gc的重组蛋白的蛋白质主链和唾液酸Neu5Gc聚糖部分的免疫原性。
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