抗CD20人鼠嵌合单抗N糖的毛细管电泳分析  

Analysis of N-linked glycan profile of human/murine chimeric anti-CD20 antibodies by capillary electrophoresis

作　　者：王文波[1] 王兰[1] 王馨[1] 于传飞[1] 张峰[1] 刘春雨[1] 陈伟[1] 李萌[1] 高凯[1] 

机构地区：[1]中国食品药品检定研究院单克隆抗体产品室,卫生部生物技术产品检定及标准化重点实验室,北京100050

出　　处：《中国新药杂志》2015年第20期2312-2316,2345,共6页Chinese Journal of New Drugs

基　　金：国家“重大新药创制”科技重大专项(2014ZX09304311-001,2012ZX09304010);中国食品药品检定研究院中青年发展研究基金资助项目(2013NB1)

摘　　要：目的:利用毛细管电泳对原研抗CD20人鼠嵌合单抗和4个生物类似药的N糖谱进行分析。方法:对单抗Fc上的N糖链利用糖苷酶F进行酶切释放,然后用8-氨基芘-1,3,6-三磺酸(APTS)染料进行标记,通过毛细管电泳对N糖链进行分析。结果:7批原研CD20单抗N糖糖型和比例基本一致。4个生物类似药虽然在糖型上与原研单抗一致,但在比例上存在不同程度的差异。SBP-C4中G0F比例显著高于原研和其他生物类似药,而G1F和G2F比例显著较低。SBP-C1,C2和C3中分别含半乳糖修饰和岩藻糖修饰的糖型比例在(原研单抗±3SD)范围内,SBP-C4半乳糖修饰糖型比例明显较低,岩藻糖修饰的糖型比例较原研单抗略高。SBP-C1和C2的CDC活性与原研单抗(RBP-1)基本一致,SBP-C3的CDC活性略高于原研单抗,SBP-C4的CDC活性显著较低。4个生物类似药候选药物的ADCC活性与原研单抗(RBP-1)基本一致。结论:在对单抗的质量控制中,尤其是对于具有Fc功能的单抗,应重视N糖链检测的重要性。Objective： To analyze the difference of N-glycan profile between original human-murine chimeric anti-CD20 antibody and four candidates of similar biotherapeutic products（ SBP）. Methods： The N-linked glycan on Fc region was released by PNGase F digestion and labeled by 8-aminopyrene-1,3,6-trisulfonic acid（ APTS）. Then,labeled glycans was analyzed by capillary electrophoresis with laser-induced fluorescence detection（ CE-LIF）. Results： Seven batches of reference biotherapeutic products（ RBPs） showed the same pattern in N linked glycosylation,and tiny difference existed in G0 / G1 percentage. The percentage of six main glycans of four SBP candidates was different from RBPs. The G0 F percentage of SBP-C4 was much higher than the RBPs and other three SBPs,and G1 F / G2 F percentage was much lower. The total percentage of N glycans that contain galactosyl /fucosly residue was similar between RBP and SBP-C1,C2 and C3. However,percentage of galactosylated glycans in SBP-C4 was much lower but percentage of fucosylated glycans was a littile higher. SBP-C1 and C2 showed a similar CDC potency with RBP-1,while SBP-C3 showed higher CDC potency than RBP-1 but SBP-C4 was much low-er. The ADCC potency between RBP-1 and four SBP candidates was similar. Conclusion： The analysis of N-glycan profile,especially in antibodies that have an Fc effector function,should be paid more attention.

关 键 词：单克隆抗体 N糖基化修饰 毛细管电泳 生物类似药 

分 类 号：R965[医药卫生—药理学]