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| Effect of Different Additive Factors on the Growth and Antibody Secretion of Hybridoma Cells |
| ZHANG Song-Yu, ZENG Xin-Yi*, ZHU Yuan-Tao*, LI Zhi-Yuan, SU Jun**, LI Qiu-Yan** |
| College of Biological Sciences/State Key Laboratory of Animal Biotech Breeding/National Demonstration Center for Experimental Education of Life Science, China Agricultural University, Beijing 100193, China |
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Abstract The hybridoma cell technique is a crucial method for monoclonal antibody production. Monoclonal antibody-producing hybridoma cells against Porcine epidemic diarrhea virus (PEDV) have been prepared in this research group. In this study, the hybridoma cell culture system was optimized to improve the efficiency of antibody secretion from hybridoma cells by adding different concentrations of additive factors, including glutamine (0~10 mmol/L), antimicrobial peptides (0~20 μg/mL), peptides (0~20 ng/mL), human epidermal growth factor (hEGF) (0~20 ng/mL), basic fibroblast growth factor (bFGF)(0~20 ng/mL), and vitamin C (VitC)(0~20 ng/mL). These factors were introduced into the hybridoma cell culture system to investigate their impact on the growth and antibody secretion of hybridoma cells by plotting growth curves and using enzyme-linked immunosorbent assay (ELISA), respectively. The results showed that the peak of cell density was reached on the 5th day when glutamine and antimicrobial peptides were added, and the maximum cell density was achieved at the addition concentration of 6 mmol/L glutamine and 10 μg/mL antimicrobial peptides. The peak of cell density was reached on the 4th day when hEGF, bFGF and VitC were added, and the maximum cell density was achieved at the addition concentration of 10 ng/mL hEGF, 20 ng/mL bFGF and 20 ng/mL VitC. The results of antibody titer detection showed that the highest antibody titer (1∶8000) was obtained in the glutamine addition group at a concentration of 6 mmol/L, the highest antibody titer (1∶16000) was obtained in the antimicrobial peptide addition group at a concentration of 10 μg/mL, the highest antibody titer (1∶16000) was obtained in the hEGF and bFGF addition groups at concentrations of 5 and 10 ng/mL, respectively, and the highest antibody titer (1∶8000) was obtained in the VitC addition group at a concentration of 10 ng/mL. In conclusion, the growth and antibody production capabilities of hybridoma cells have been effectively enhanced through the modification of medium components. This study establishes a solid groundwork for the formulation of specialized hybridoma cell culture media aimed at production purposes.
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Received: 05 March 2024
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Corresponding Authors:
** su_jun@cau.edu.cn; liqiuyan-yuan@163.com
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| About author:: * These authors contributed equally to this work |
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