Lyon Robert P, Meyer David L, Setter Jocelyn R, Senter Peter D
Seattle Genetics, Bothell, Washington, USA.
Methods Enzymol. 2012;502:123-38. doi: 10.1016/B978-0-12-416039-2.00006-9.
Many methods have been described for the conjugation of drugs to monoclonal antibodies. The presence of a discrete number of readily reducible disulfides in the common IgG subtypes presents a convenient opportunity for conjugation to cysteine residues with thiol-reactive drug-linkers. Such conjugates can be prepared by a straightforward two-step reaction scheme involving the reduction of the antibody disulfides to the desired number of average thiols per antibody, followed by addition of the drug-linker, ideally with a maleimido functionality for rapid, selective reaction. In a discovery setting, this basic method can be scaled down to produce microgram quantities of conjugate for early screening, and in a manufacturing setting can be scaled up to produce grams or kilograms of conjugate for clinical trials and commercialization. The resulting conjugates are readily characterized using common HPLC methods.
已经描述了许多将药物与单克隆抗体偶联的方法。常见IgG亚型中存在数量离散的易于还原的二硫键，这为与具有硫醇反应性药物连接体的半胱氨酸残基偶联提供了便利的机会。此类偶联物可通过一个简单的两步反应方案制备，该方案包括将抗体二硫键还原为每个抗体所需数量的平均硫醇，然后加入药物连接体，理想情况下该连接体具有马来酰亚胺官能团以实现快速、选择性反应。在发现阶段，这种基本方法可以缩小规模以生产微克量的偶联物用于早期筛选，而在生产阶段则可以扩大规模以生产克或千克量的偶联物用于临床试验和商业化。使用常见的高效液相色谱方法可以很容易地表征所得的偶联物。
