Center for Molecular Immunology, Atabey Siboney, Playa, Havana 11600, Cuba.
BMC Biotechnol. 2011 Nov 22;11:112. doi: 10.1186/1472-6750-11-112.
1E10 monoclonal antibody is a murine anti-idiotypic antibody that mimics N-glycolyl-GM3 gangliosides. This antibody has been tested as an anti-idiotypic cancer vaccine, adjuvated in Al(OH)3, in several clinical trials for melanoma, breast, and lung cancer. During early clinical development this mAb was obtained in vivo from mice ascites fluid. Currently, the production process of 1E10 is being transferred from the in vivo to a bioreactor-based method.
Here, we present a comprehensive molecular and immunological characterization of 1E10 produced by the two different production processes in order to determine the impact of the manufacturing process in vaccine performance. We observed differences in glycosylation pattern, charge heterogeneity and structural stability between in vivo-produced 1E10 and bioreactor-obtained 1E10. Interestingly, these modifications had no significant impact on the immune responses elicited in two different animal models.
Changes in 1E10 primary structure like glycosylation; asparagine deamidation and oxidation affected 1E10 structural stability but did not affect the immune response elicited in mice and chickens when compared to 1E10 produced in mice.
1E10 单克隆抗体是一种模拟 N-糖基酰基-GM3 神经节苷脂的鼠抗独特型抗体。该抗体已作为抗独特型癌症疫苗进行测试，与 Al(OH)3 联合使用，在黑色素瘤、乳腺癌和肺癌的几项临床试验中进行了测试。在早期临床开发中，这种 mAb 是从小鼠腹水液中体内获得的。目前，1E10 的生产过程正在从体内向基于生物反应器的方法转移。
在这里，我们对通过两种不同生产工艺生产的 1E10 进行了全面的分子和免疫学表征，以确定生产工艺对疫苗性能的影响。我们观察到体内产生的 1E10 和生物反应器获得的 1E10 在糖基化模式、电荷异质性和结构稳定性方面存在差异。有趣的是，这些修饰对两种不同动物模型中引起的免疫反应没有显著影响。
与在小鼠中产生的 1E10 相比，1E10 一级结构的变化（如糖基化；天冬酰胺脱酰胺和氧化）影响了 1E10 的结构稳定性，但并未影响在小鼠和鸡中引起的免疫反应。
