Study On Serum-Free Culture Technique Of Anti-Ferritin Hybridoma Cells

Hybridoma cells have the dual advantages of unlimited reproduction and secretion of antibodies,and their large-scale culture process is an important technology for the production of antibody products.Large-scale serum-free culture of hybridoma cells has the advantages of simplifying the process,reducing production costs,and increasing antibody production.In this paper,the effects of serum on myeloma and hybridoma cells were studied.Based on this,a serum-free medium suitable for the growth of hybridoma cells was developed,and the expansion culture of the serum-free medium of hybridoma cells was further studied.The main content includes the following parts:Firstly,DMEM medium containing different serum concentrations(10%,7.5%,5%)was prepared,and the three cells with good growth status,log phase 5H2,Sp2/0-Ab,and Sp2/0were treated with three different kinds.The inoculation concentration was inoculated into a12-well cell culture plate containing the above medium.The effects of different serum concentrations and different seeding densities on cell growth and proliferation were examined.The data measured by the cytometer showed that the serum concentration had a great influence on the three cells.The three cells grew normally in the 10%serum concentration medium and could not grow normally in the low serum concentration.At the same serum concentration,the cells were Cells with high seeding density proliferate faster and have a shorter doubling time.Secondly,the DMEM medium,Ham's F-12 medium and RPMI1640 medium were formulated into a basic medium according to a certain ratio.On this basis,different amounts of insulin,transferrin,cholesterol and ethanolamine were added to form a serum-free low-protein medium.The growth of the cells in the serum-free medium and the commercially available serum-free medium was examined by serum-free acclimation of the cells,and it was revealed that the prepared Z4 medium had the same effect as the commercially available medium,and the cells could grow normally.Thirdly,the 5H2 cell spinner flask culture was carried out separately with Z4 and H-SFM medium,and the changes of glucose,glutamine,lactic acid ammoina and antibody concentrations in the medium were measured.The results showed that the number of cells in the H-SFM medium reached a peak of 14.57×10~5 cells/mL on the fifth day.The rate of antibody production was significantly increased,and the antibody concentration was 39?g/mL on the seventh day.The number of cells in the Z4 medium reached a peak of19.87×10~5 cells/mL on the sixth day,and the antibody concentration on the seventh day was56?g/mL.Both glucose and glutamine concentrations in Z4 medium and H-SFM medium showed a downward trend,and the concentration of lactic acid showed an upward trend.Appropriate increase in the initial concentration of glucose and glutamine also increased cell growth.