Profiling Antibody Drug Conjugate Positional Isomers: A System-of-Equations Approach
Profiling Antibody Drug Conjugate Positional Isomers: A System-of-Equations Approach
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DOI：
10.1021/ac301568f
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发表时间：
2012-09-04
影响因子：
7.4
通讯作者：
Galush, William J.
中科院分区：
文献类型：
--
作者：
Le, Lan N.;Moore, Jamie M. R.;Galush, William J.
抗体药物偶联物能够将有效的化疗剂直接靶向递送至癌细胞。它们是通过细胞毒素与单克隆抗体的化学偶联制备的，这可以通过首先还原链间二硫键，然后将所得游离巯基与药物偶联来实现。该过程产生受控但异质的缀合产物群体，其含有具有与抗体上的不同前链间二硫键半胱氨酸残基连接的不同数量的药物的物质。我们已经开发了一种数学方法，使用毛细管电泳和疏水相互作用色谱的输入来确定抗体药物缀合物群体内的位置异构体分布。通过分析特定药物-抗体比物种的分离样品确认结果。该方法适用于位置异构体分布的快速测定，并且具有材料要求低的特点。基于相同IgG框架和小分子药物组合的几种抗体药物缀合物的调查显示，使用该技术，异构体在所有分子中的分布非常相似，表明了稳健的缀合过程。
Antibody drug conjugates enable the targeted delivery of potent chemotherapeutic agents directly to cancerous cells. They are made by the chemical conjugation of cytotoxins to monoclonal antibodies, which can be achieved by first reducing interchain disulfide bonds followed by conjugation of the resulting free thiols with drugs. This process yields a controlled, but heterogeneous, population of conjugated products that contains species with various numbers of drugs linked to different former interchain disulfide cysteine residues on the antibodies. We have developed a mathematical approach using inputs from capillary electrophoresis and hydrophobic interaction chromatography to determine the positional isomer distribution within a population of antibody drug conjugates. The results are confirmed by analyzing isolated samples of specific drug-to-antibody ratio species. The procedure is amenable to rapid determination of positional isomer distributions and features low material requirements. A survey of several antibody drug conjugates based on the same IgG framework and small molecule drug combination has shown a very similar distribution of isomers among all of the molecules using this technique, suggesting a robust conjugation process.