重组抗EGFR人鼠嵌合单克隆抗体原液生产车间工艺设计.docxVIP

PAGE 17 PAGE 1 重组抗EGFR人鼠嵌合单克隆抗体原液生产车间工艺设计 摘 要 随着单抗药物的日益发展，现如今单抗药物的生产能力远不能满足市场的需求，具有鼠源性的单克隆抗体不适合用于人体，因其用于人体会引起HAMA反应，即人抗鼠抗体反应；且通过杂交瘤技术制备而成的单克隆抗体，是鼠源性的。解决此难题的方法在于可将鼠源抗体的可变区，与人抗体的恒定区进行拼接，两者拼接而形成嵌合抗体；另外，鼠抗体内可变区的互补决定区，即CDR区与人的抗体内的互补决定区，可以进行互换，两者互换可构成人源化抗体，这也是解决HAMA反应的一种方法。 至今为止，国内外已经上市的抗EGFR单克隆抗体，共有3种，将其与其他的化疗药进行比较，具备许多特点，首先其靶点非常明确，其次其毒副作用也较低，且疗效更为明显。本次设计的主要内容是设计生产西妥昔单抗的车间，主要分为上游动物细胞发酵培养和下游产物纯化两个部分，本设计选用的宿主细胞，是表达重组抗EGFR人鼠嵌合单克隆抗体的CHO细胞，对其进行悬浮培养，以分批流加补料方式培养，下游采用亲和层析、离子交换层析等对产物进行纯化。按照所有的工艺流程对所用物料进行物料衡算。原液车间内容包括：种子复苏、种子扩大培养、发酵罐发酵、蛋白纯化几个步骤。 关键词：动物细胞培养；纯化；物料衡算；车间设计 Process design of workshop for production of recombinant human mouse chimeric monoclonal antibody against EGFR Abstract Today, with the development of single drug resistant capacity of the single drug resistant far cannot satisfy the demand of the market, and because by hybridoma technology preparation of monoclonal antibody is the source of the rat, apply to the human body inevitably cause anti mouse antibody (HAMA) response, so will be the source of the rat antibodies variable area with constant region joining together to form a chimeric antibody or will be decided the mouse antibody variable area complementary area (CDR) and antibody of complementary decision of the district swap humanized antibody, basically settled the problem of antibody drugs of different source sex. At present, there are three kinds of anti-egfr monoclonal antibodies on the market at home and abroad. Compared with other chemotherapeutic drugs, they have clear targets, significant efficacy and low toxic and side effects, and have achieved good clinical efficacy. The product was approved for domestic market in 2009. This design mainly aims at the workshop production of cetuximab to carry on the design, the main product is divided into animal cells fermentation culture and downstream purification two parts, choose express recombinant EGFR resistant human mouse chimeric monoclonal antibody of CHO cells as a host cell, suspension culture an