Discovery of a chemical modification by citric acid in a recombinant monoclonal antibody.
Discovery of a chemical modification by citric acid in a recombinant monoclonal antibody.
复制标题
DOI：
10.1021/ac502179m
复制
发表时间：
2014-09-16
影响因子：
7.4
通讯作者：
Zhou, Zhaohui Sunny
中科院分区：
文献类型：
--
作者：
Chumsae, Chris;Zhou, Liqiang Lisa;Shen, Yang;Wohlgemuth, Jessica;Fung, Emma;Burton, Randall;Radziejewski, Czeslaw;Zhou, Zhaohui Sunny
重组治疗性单抗表现出高度的异质性，这种异质性可以由各种翻译后修饰产生。蛋白质产品的配方是保持特定的pH值，并尽量减少进一步的修改。柠檬酸通常被认为是安全的(GRAS)，通常用于将PH值保持在3到6之间的配方，通常被认为是化学惰性的。然而，正如我们在此报道的，柠檬酸在pH 5.2的磷酸盐/柠檬酸盐缓冲配方中共价修饰重组单抗(IgG1)，并导致所谓的“酸性物种”的形成，其质量分别增加了174和156 Da。肽图谱显示，修饰发生在轻链的N-末端。另外三种抗体也表现出相同的修饰，但对轻链、重链或两者的N-末端显示不同的敏感性。因此，在某些条件下，表面上无反应的辅料可能会增加配制的重组单抗中的异质性和酸性物种。以此类推，其他具有两个或更多羧酸基团并能够形成酸酐的分子(例如琥珀酸)也可以表现出类似的反应活性。总之，我们的发现再次提醒我们，将配方视为化学修饰和产品异质性的潜在来源是谨慎的。
Recombinant therapeutic monoclonal antibodies exhibit a high degree of heterogeneity that can arise from various post-translational modifications. The formulation for a protein product is to maintain a specific pH and to minimize further modifications. Generally Recognized as Safe (GRAS), citric acid is commonly used for formulation to maintain a pH at a range between 3 and 6 and is generally considered chemically inert. However, as we reported herein, citric acid covalently modified a recombinant monoclonal antibody (IgG1) in a phosphate/citrate-buffered formulation at pH 5.2 and led to the formation of so-called “acidic species” that showed mass increases of 174 and 156 Da, respectively. Peptide mapping revealed that the modification occurred at the N-terminus of the light chain. Three additional antibodies also showed the same modification but displayed different susceptibilities of the N-termini of the light chain, heavy chain, or both. Thus, ostensibly unreactive excipients under certain conditions may increase heterogeneity and acidic species in formulated recombinant monoclonal antibodies. By analogy, other molecules (e.g., succinic acid) with two or more carboxylic acid groups and capable of forming an anhydride may exhibit similar reactivities. Altogether, our findings again reminded us that it is prudent to consider formulations as a potential source for chemical modifications and product heterogeneity.
登录
查看更多内容
影响因子：
4.8
作者：
Chen, Tingsu;Nayak, Nihar;Downie, A. Bruce
通讯作者：
Downie, A. Bruce
DOI：
10.1016/j.jasms.2008.11.011
发表时间：
2009-03-01
影响因子：
3.2
作者：
Liu, Hongcheng;Gaza-Bulseco, Georgeen;Zhou, Lisa
通讯作者：
Zhou, Lisa
影响因子：
15
作者：
HIGUCHI, T;MIKI, T;HERD, AK
通讯作者：
HERD, AK
DOI：
10.1016/j.jchromb.2004.12.033
发表时间：
2005-04-25
影响因子：
3
作者：
Cordoba, AJ;Shyong, BJ;Harris, RJ
通讯作者：
Harris, RJ
DOI：
10.1016/j.jchromb.2008.10.015
发表时间：
2008-12-01
影响因子：
3
作者：
Liu, Hongcheng;Gaza-Bulseco, Georgeen;Lundell, Edwin
通讯作者：
Lundell, Edwin