Using Six Sigma Methods To Research The Stability Of Yields Of Antibody Production By Animal Cells

Since antibody drug can cure many incurable diseases, it has highly attracted the attention of bio-phannaceutical industry. Currently, the use of large-scale animal cell culture technology to produce genetically engineered antibody has become mainstream,but the stability of antibody has restricted its further development. To solve this problem, Chinese hamster ovary cells producing antibodies process were studied in this article. Six Sigma management methods were used to regulate critical parameters of antibody production, and to make the process of antibody production a statistically stable one. This study can provide technical support to bio-pharmaceutical companies to ensure the stability of its products.It was found that under the same conditions, the yields of antibody obtained from different batches were quite different. To determine the critical factors of antibody production process, cell line c was studied. Failure mode and effects analysis method was used to identify and assess the risk of antibody production process. First, professional team was established and listed all the factors of the antibody production process in the form of fish bone diagram. Then convert the fish bone diagram into a questionnaire. Each item of questionnaire should be evaluated and scored in three aspects, which are severity, occurrence and detection. After that, risk priority number of each item was obtained and was ranked from the top to the bottom. Error of measuring instrument, cell specific growth rate, initial pH of basal medium, sample volume or the like were screened to be the potential critical process parameters in the process of antibody production.Since the error of measuring instrument receives the highest risk priority number by failure mode and effects analysis, measurement statistical method of Six Sigma was used to measure main instruments of antibody production process, including high performance liquid chromatography,10 mL pipettes,1 mL pipette, cell count boards and pH meter. Bias, linearity, repeatability and process capability were studied by measurement system analysis. It turns out that bias and linearity of HPLC, pH meter,10 mL pipette and 1 mL pipette are within the acceptable range, so the accuracy is eligibe; Repeatability and resolution of HPLC, pH meter, blood cell count reached procedure requirement; through process capability analysis, the process capability of 10 mL pipette and 1 mL pipette was acceptable. These test results of measuring instruments after modifications are in line with the needs of the process.Factorial experiment and single factor experiments were used to analyze the rest high-risk factors.It was found that the seed cell specific growth rate, replacement of N-1 generation seed medium, initial pH value of basal medium, sample volume, standing time after centrifugation in centrifugal inoculation process and the initial inoculation cell density were significant factors affecting yield of antibody. Accordingly, original production process of the cell line c was supplemented and improved. Statistical process analysis showed that the stability of the new process of antibody production has been improved.The results of this study can provide a reference for the antibody-producing industries to improve yield and quality of antibody drugs, to reduce the long-term production costs and to improve the ideas of production.