CORE--LARGE SCALE CULTURE AND PURIFICATION

该计划项目的目标是证明无线电标记的、 基因工程抗体可用于治疗人类结肠癌。这个 核心9003的责任，大规模培养和提纯 抗体，是生产足够数量的工程抗体和 人体成像和I/II期治疗研究的纯度 计划的重点，以及项目1和6中的研究研究。 据估计，最初的第一阶段试验将需要生产 提纯2.4-3.0克cT84.66。核心将协助 项目2(工程抗体)用于鉴定高产菌株 每种基因工程抗体的细胞系。这些细胞系将 根据美国食品及药物管理局的规定进行测试 用于生产生物制品的细胞系的特征，(1987)“，和 在制备和检测单抗时应注意的几个问题 人用产品，(1987)“。抗体将在以下状态下产生- 最先进的中空纤维生物反应器，能够生产革兰氏 抗体量，既浓缩又相当纯净 当它被收获的时候。该抗体将通过蛋白G亲和纯化。 阴离子交换层析。纯化的抗体将满足 FDA中规定的规范“是指 人用单抗制品的研制和检测， (1987)“。将特别注意以下要求： 纯化的抗体通过了适当的热原测试，它含有 &lt；每针10 pg DNA。纯化的抗体将被转移 到核心9001(放射性药物)，在那里它将与一个 适当的螯合剂和放射性标记，以供后续的人体试验。

The goal of this Program Project is to demonstrate that radiolabeled, engineered antibodies can be used to treat human colonic cancer. The responsibility of Core 9003, Large-Scale Culture and Purification of Antibody, is to produce engineered antibody of sufficient quantity and purity for the human imaging and Phase I/II therapy studies that are focus of the program, and for the research studies in Projects 1 and 6. It is estimated that the initial Phase I trial will require production and purification of 2.4-3.0 grams of cT84.66. The Core will assist Project 2 (Engineered Antibodies) in the identification of high-producing cell lines for each of the engineered antibodies. These cell lines will be tested in accordance with the FDA "Points to Consider in the Characterization of Cell Lines Used to Produce Biologicals, (1987)", and "Points to Consider in the Manufacture and Testing of Monoclonal Antibody Products for Human Use, (1987)". Antibody will be produced in a state- of-the-art hollow-fiber bioreactor which is capable of producing gram quantities of antibody, which is both concentrated and reasonably pure as it is harvested. The antibody will be purified by Protein G affinity and anion exchange chromatography. The purified antibody will meet the specifications set forth in the FDA "Points to Consider in the Manufacture and Testing of Monoclonal Antibody Products for Human Use, (1987)". Specific attention will be given to requirements that the purified antibody pass an appropriate pyrogen test and that it contain < 10 pg DNA per injectable dose. Purified antibody will be transferred to Core 9001 (Radiopharmacy) where it will be conjugated with an appropriate chelator and radiolabeled for subsequent human trials.