Ye X.,Lanzhou Institute of Biological Product Co. |
Mao X.-Y.,Lanzhou Institute of Biological Product Co.
Chinese Journal of Biologicals | Year: 2016
Recombinant monoclonal antibody is one of the hot spots in recent years because of its high specificity and significantly curative effect, which is of a wide prospect in the treatment of tumor, auto-immune diseases and infectious diseases as well as transplant rejection reactions. Since antibody therapy requires large doses over a long period of time, the mammalian cell (particularly Chinese hamster ovary cells) culture based on bioreactors have become the core technology of the large scale manufacturing of monoclonal antibodies. The techniques for culture of mammalian include the screening of engineered cell line, development of optimization of serum-free media, as well as optimization of culture process in bioreactor. This article focuses on the development, classification and optimization strategy of serum-free media, so as to provide a reference for development of economic and effective production process for monoclonal antibody drags.
Chang Y.-L.,Lanzhou Institute of Biological Product Co.
Chinese Journal of Biologicals | Year: 2013
Objective: To develop and verify a double antigen sandwich ELISA kit for Yersinia pestis F1 antibody. Methods: The double antigen sandwich ELISA kit for Y. pestis F1 antibody was developed using purified F1 antigen as coating antigen and HRP-labeled F1 antigen as enzyme labeled antigen. Three batches of kits were prepared, and internal reference was set up. The kit was verified for precisions in intra- and inter-assays, sensitivity, specificity, accuracy and stability. Results: The optimal concentration of F1 antigen for coating was 0.6 μg/ml, while the optimal working concentration of HRP-labeled F1 antigen was 1:7 000. The CVs of results in intra- and inter-assays on reference for precision were 6.5% and 7.1% respectively. The recovery rates of internal reference and the reference for precision at high, moderate and low concentrations were 95%-112%. Both the sensitivities of the kit for positive and negative sera were 100%. No cross reactions with normal human serum, normal rabbit serum, normal mouse serum as well as rabbit and mouse sera against Y. pseudotuberculosis were observed. The kit showed high stability, of which the validity period was at least 12 months. Conclusion: The double antigen sandwich ELISA kit for Y. pestis F1 antibody was successfully developed, which met the requirements for license of in vitro diagnostic kit, and was suitable for the clinical monitoring, diagnosis and prognosis of plaque.
Ren K.-M.,Lanzhou Institute of Biological Product Co. |
Zhang Y.,Lanzhou Institute of Biological Product Co. |
Wang J.-H.,Lanzhou Institute of Biological Product Co. |
Wang X.,Lanzhou Institute of Biological Product Co. |
And 2 more authors.
Chinese Journal of Biologicals | Year: 2013
Objective: To optimize the ultrafiltration process for capsular polysaccharide of Streptococcus pneumoniae type 14 by response surface methodology. Methods: The performance of membrane was investigated with respect to three factors including transmembrane pressure (TMP), cross-flow rate and polysaccharide concentration. The effects of various factors and their interactions on the performance of membrane were also investigated by Box-Behnken Design. The predictive model of regression equation was analyzed by Design Expert software. The optimized process parameters were verified by using three batches of capsular polysaccharide of S. pneumonia type 14. Three batches of the capsular polysaccharide were subjected to equivoluminal dialysis by the optimal ultrafiltration process, based on which the end point of ultrafiltration was determined. Overall control tests were performed on the purified capsular polysaccharide. Results: The optimal TMP, cross-flow rate, polysaccharide concentration and diavolume for ultrafiltration were 12.8 Psi, 66 L/h, 0.86 g/L and 5 respectively. Under such a condition, the predictive performance of membrane was 91.7 g/(h·m2), while the actual value was 91.9 g/(h·m2). All the polysaccharide components in three batches of capsular polysaccharide of S. pneumonia type 14 met the relevant requirements in European Pharmacopoeia (7. 0 edition), while the relative molecular masses were more than 420 000, and no residual sodium deoxycholate or ethanol was detected. The three batches of capsular polysaccharide showed good antigenicity, of which the antigenic components showed no difference. Conclusion: By using the ultrafiltration process for capsular polysaccharide of S. pneumonia type 14 optimized by response surface methodology, the working efficiency increased significantly, while the production cost decreased, which provided an experimental data for large-scale production of the capsular polysaccharide.