CORE--LARGE SCALE CULTURE AND PURIFICATION

本计划项目的目标是证明放射性标记， 工程抗体可用于治疗人结肠癌。 的 核心9003的责任，大规模培养和纯化 抗体，是生产足够数量的工程抗体， 用于人体成像和I/II期治疗研究的纯度， 该计划的重点，并在项目1和6的研究。 据估计，第一阶段试验将需要生产 和纯化2.4-3.0克cT84.66。 核心会协助 项目2（工程抗体）在鉴定高产 每种工程化抗体的细胞系。 这些细胞系将 根据FDA的“考虑要点 用于生产生物制品的细胞系的表征，（1987）"，以及 “单克隆抗体生产和检测中的考虑要点 人用产品（1987年）"。 抗体将在一种状态下产生- 现有技术的中空纤维生物反应器能够生产克 一定量的抗体，其既浓缩又相当纯 当它被收获的时候。 抗体将通过蛋白G亲和纯化 和阴离子交换色谱法。 纯化的抗体将满足 FDA“在使用过程中应考虑的要点”中规定的规范 人用单克隆抗体产品的生产和检测， （1987年）"。 将特别注意以下要求： 纯化的抗体通过适当的热原试验，并且含有 < 10 pg DNA/注射剂量。 将纯化抗体转移至 核心9001（放射性药物学），其中它将与 合适的螯合剂并放射性标记用于随后的人体试验。

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.