Preparation Process And Quality Research For Bevacizumab

Bevacizumab used in curing some kinds of cancer is the first drug of inhibiting tumor angiogenesis approved by USA FDA. The aim of the subject is to research and develop the preparation process of Bevacizumab, and then the quality study is to identify the structure and quality attribute and stability. All study work is the basis for commercialized production. The ultimate aim is to manufacture Bevacizumab that can be affordable for Chinese people and to improve life quality of patients.The light-chain and heavy-chain genes of bevacizumab are cloned by recombinant DNA technology. Insert two genes chain into retrovirus front vector of high-efficiency expression.And the cells 293 GP are transfected by light chain and heavy-chain genes expression front vector with virus plasmid p HCMV-G separately. Obtain two kinds of retrovirus vectors with light-chain and heavy-chain genes of bevacizumab. The two vectors are transduced successively into cell CHO-S to get the engineering cell expressed bevacizumab stably. Amplify the cells through advanced cell culture technology and express bevacizumab. Purify the bevacizumab through process including inactivation and remove of virus. And then obtain to finished products through some formulation manufacturing process including preparation, filtrating and sterility filling process,etc.Grope and optimize the process parameter of cell culture and purification by several tests to confirm the manufacturing process high-efficiency expression for bevacizumab. Meanwhile, bulk solution and finished products manufactured by the process are analyzed by quality study and stability study. All test results are conformed. Engineering cell passage is good stability. The cell expression is higher. Purification process can obtain high assay of bulk solution. Batch-to-Batch quality is consistent by quality research for seven batches of bulk solution and finished products. Primary structure of protein manufactured by the process is the same as reference AVASTIN. The formulation of finished product is better than that of AVASTINâ—‹R. Batch-to-Batch quality and structure is consistent with reference AVASTINâ—‹R by identifying the structure and quality attribute for three batches( One batch used in N- Sugar spectrum analysis, glycosylation site analysis and disulfide bonding analysis.The manufacturing process researched in the subject is achievable for preparation of bevacizumab and manufacture the products resembling with RLD stably and continuously. The manufacturing process can be applied to large-scale production to perform the clinical research.