HPMA copolymer-anticancer drug-OV-TL16 antibody conjugates .1. Influence of the method of synthesis on the binding affinity to OVCAR-3 ovarian carcinoma cells in vitro
HPMA copolymer-anticancer drug-OV-TL16 antibody conjugates .1. Influence of the method of synthesis on the binding affinity to OVCAR-3 ovarian carcinoma cells in vitro
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DOI：
10.3109/10611869608996827
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发表时间：
1996-01-01
影响因子：
4.5
通讯作者：
Kopecek, J
中科院分区：
文献类型：
--
作者：
Omelyanenko, V;Kopeckova, P;Kopecek, J
研究了将 OV-TL16 抗体及其 Fab' 片段与 N-(2-羟丙基)甲基丙烯酰胺 (HPMA) 共聚物-药物（阿霉素 {ADR} 或内消旋二氯林 e(6) 单（N-2-氨基乙基酰胺）{Mce(6)}）缀合物结合的不同方法对缀合物与卵巢癌 (OVCAR-3) 细胞相关抗原的亲和力的影响。抗体通过氨基与 HPMA 共聚物-药物（ADR 或 Mce(6)）缀合物结合，产生抗原结合异质的缀合物。将 HPMA 共聚物-Mce(6) 缀合物与抗体的碳水化合物区域偶联，产生的缀合物比通过氨基将抗体与聚合物连接而制备的缀合物具有更均匀的亲和常数分布。然而，与天然抗体相比，这两种方法都会导致亲和常数降低。用 OV-TL16 抗体的 Fab' 片段制备的缀合物表现出比用完整抗体制备的任一缀合物更均匀的亲和力。为了验证结合亲和力和均质性的变化是抗体结构构象变化的结果这一假设，采用了一系列物理化学方法来表征缀合物。 OV-TL16抗体和药物（ADR和Mce(6)）之间的激发能量转移和Mce(6)的光谱特性用于监测抗体和药物之间的相互作用。抗体固有荧光的猝灭也用于研究其构象变化。已尝试将细胞表面的生物识别与药物与抗体分子的相互作用以及抗体构象的变化联系起来。
The influence of different methods of binding the OV-TL16 antibody and its Fab' fragment to N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer-drug (adriamycin {ADR} or meso chlorin e(6) mono(N-2-aminoethylamide) {Mce(6)}) conjugates on the affinity of conjugates to an ovarian carcinoma (OVCAR-3) cell associated antigen was investigated. The binding of the antibody to HPMA copolymer-drug (ADR or Mce(6)) conjugates via amino groups resulted in conjugates which were heterogeneous in their antigen binding. Coupling the HPMA copolymer-Mce(6) conjugate to the carbohydrate region of the antibody resulted in conjugates with a more homogeneous distribution of affinity constants than conjugates prepared by linking the antibody to the polymer via amino groups. However, both methods resulted in a decrease in the affinity constant compared to the native antibody. Conjugates prepared with the Fab' fragment of the OV-TL16 antibody demonstrated a more homogeneous affinity than either conjugate prepared with the whole antibody. To verify the hypothesis that the changes in the binding affinity and homogeneity are a consequence of conformational changes in the antibody structure, a series of physicochemical methods were employed to characterize the conjugates. The excitation energy transfer between OV-TL16 antibody and drugs (ADR and Mce(6)) and the spectral properties of Mce(6) were used to monitor the interactions between the antibody and drugs. The quenching of the intrinsic fluorescence of the antibody was also employed to study its conformational changes. An attempt has been made to correlate the biorecognition at the cellular surface with the interactions of drug with the antibody molecule and with the changes in antibody conformation.