Jiang F N, Jiang S, Liu D, Richter A, Levy J G
Department of Microbiology, University of British Columbia, Vancouver, Canada.
J Immunol Methods. 1990 Nov 6;134(1):139-49. doi: 10.1016/0022-1759(90)90122-c.
A procedure is described whereby the photosensitizer, benzoporphyrin derivative monoacid ring A (BPD-MA) was covalently linked to a model monoclonal antibody in a manner which is reproducible, quantifiable, and retains both the biological activity of the antibody and the cytotoxicity of the photosensitizer. Preliminary steps involved the linkage of BPD-MA to a modified polyvinyl alcohol (PVA) backbone, followed by conjugation to the antibody using heterobifunctional linking technology. Briefly, polyvinyl alcohol (MW ca. 10,000) was modified with 2-fluoro-1-methyl pyridinium toluene-4-sulfonate and 1,6-hexanediamine to produce side chains containing free amino groups. The free carboxyl group of BPD-MA was utilized to conjugate photosensitizer molecules to modified PVA using a standard carbodiimide reaction. Final linkage of the PVA-BPD to a model monoclonal antibody involved further substitution of the carrier with 3-mercaptopropionic acid and carbodiimide to introduce 3-4 sulfhydryl residues per carrier molecule, and introduction of sulfo-m-maleimidobenzoyl-N-hydroxysulfosuccinimide ester residues to the monoclonal (3-4 residues/molecule). Conjugation was effected by reaction of the two species at pH 5.5 for 18 h. Detailed methodology and tests for efficacy of the procedure are provided.
本文描述了一种方法，通过该方法将光敏剂苯并卟啉衍生物单酸环A（BPD-MA）以可重复、可量化的方式共价连接到模型单克隆抗体上，同时保留抗体的生物活性和光敏剂的细胞毒性。初步步骤包括将BPD-MA连接到修饰的聚乙烯醇（PVA）主链上，然后使用异双功能连接技术将其与抗体偶联。简要地说，用对甲苯磺酸2-氟-1-甲基吡啶鎓和1,6-己二胺对聚乙烯醇（分子量约10,000）进行修饰，以产生含有游离氨基的侧链。利用BPD-MA的游离羧基，通过标准的碳二亚胺反应将光敏剂分子与修饰的PVA偶联。PVA-BPD与模型单克隆抗体的最终连接涉及用3-巯基丙酸和碳二亚胺进一步取代载体，以使每个载体分子引入3-4个巯基残基，并将磺基间马来酰亚胺苯甲酰-N-羟基磺基琥珀酰亚胺酯残基引入单克隆抗体（每个分子3-4个残基）。通过使两种物质在pH 5.5下反应18小时来实现偶联。文中提供了该方法的详细步骤和效果测试。
