Matsuda Yutaka, Chakrabarti Atis, Takahashi Kazutoshi, Yamada Kei, Nakata Kunio, Okuzumi Tatsuya, Mendelsohn Brian A
Ajinomoto Co., Inc., 1-1 Suzuki-cho, Kawasaki, Kanagawa 210-8681, Japan.
Tosoh Bioscience, 3604 Horizon Drive, Suite 100, King of Prussia, PA 19406, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2021 Jul 1;1177:122753. doi: 10.1016/j.jchromb.2021.122753. Epub 2021 May 19.
Commercially approved conventional antibody-drug conjugates (ADCs) are produced as heterogeneous mixtures containing a stochastic distribution of payloads decorating the antibody molecules resulting in decreased efficacy and thus lowering their therapeutic index. Control of the DAR and conjugation site in the development of next-generation ADCs is believed to assist in increasing the therapeutic index of these targeted biologics leading to overall enhanced clinical efficacy and reduced toxicity. A chemical site-specific conjugation technology termed AJICAP® allows ADC developers to control both the location and quantity of the payload conjugation to an antibody. Furthermore, this simplified ADC composition enables a streamlined chemical analysis. Here we report the chromatographic separation of site-specific ADCs produced by AJICAP® technology using an analytical affinity chromatography HPLC column containing a recombinant FcγIIIa receptor-ligand immobilized on a non-porous polymer resin (NPR). These HPLC analyses provided visually clear chromatogram results reflecting the heterogeneity of each ADC. The affinity strength was also measured by biolayer interferometry (BLI) and predicted by molecular structure analysis. The results indicate that AJICAP® technology is a promising solution to link hydrophobic payloads to antibodies without compromising antibody receptor function. This study also shows that FcγIIIa-NPR column can be used to characterize site-specific conjugated ADCs compared to ADCs synthesized using conventional methods.
商业上批准的传统抗体药物偶联物（ADC）是作为异质混合物生产的，其中负载物随机分布在抗体分子上，导致疗效降低，从而降低其治疗指数。据信，在下一代ADC的开发中控制药物与抗体的比率（DAR）和偶联位点有助于提高这些靶向生物制剂的治疗指数，从而全面提高临床疗效并降低毒性。一种称为AJICAP®的化学位点特异性偶联技术使ADC开发者能够控制负载物与抗体偶联的位置和数量。此外，这种简化的ADC组成使化学分析更加简化。在此，我们报告了使用含有固定在无孔聚合物树脂（NPR）上的重组FcγIIIa受体配体的分析亲和色谱HPLC柱，对通过AJICAP®技术生产的位点特异性ADC进行色谱分离。这些HPLC分析提供了直观清晰的色谱图结果，反映了每个ADC的异质性。还通过生物层干涉术（BLI）测量了亲和强度，并通过分子结构分析进行了预测。结果表明，AJICAP®技术是一种很有前景的解决方案，可将疏水性负载物与抗体连接起来，而不会损害抗体受体功能。这项研究还表明，与使用传统方法合成的ADC相比，FcγIIIa-NPR柱可用于表征位点特异性偶联的ADC。
