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After stable transfection, it is usually necessary to perform a large number of screens. Creative Biogene provides advanced technology and screening protocols for rapid selection of optimal cell pools after transfection and helps evaluate overall cloning performance in terms of protein concentration, cell health, and viability.
The plasmids transfected with host cells carry some kind of selective markers (e.g. metabolic markers, antibiotic markers, etc.). After transferring the plasmids with markers into the host cells, the selective medium is used to screen the plasmids. Host cells that are not successfully transfected with plasmids or are not at the right transfection site will die because they lack the selective markers and cannot grow on the selective medium, and positive clones that are successfully transfected can grow on the selective medium, thus achieving the purpose of screening positive clones. In the amplification selection system, a corresponding selection pressure will also be applied to the transfected host cells, and only cell lines with high levels of exogenous gene expression will survive, thus screening for stable cell lines with high expression.
Creative Biogne utilizes our advanced technology and screening strategies to screen and amplify mini-pools. Our experienced and skilled scientists disperse the transfected cultures into 96-well plates and place them under the appropriate selection conditions for screening. Each mini-pool is evaluated by advanced flow cytometry. Mini-pools with the highest level of amplification will be selected through a series of evaluations to identify the best mini-pool for expanded expression in shake flasks.
We analyze the expression of mini-pools in shake flasks using a replenishment batch process and perform periodic assessments. On each assessment day, cell viability, viable cell density and titer are measured, and retained samples are archived. We prepare super pools by combining cells from the top mini-pools that present similar titers based on titer data. The number of super pools generated and the selection of mini-pools is at the sole discretion of Creative Biogene.
Generating high-yield cell lines is resource intensive and often requires a long timeline and screening of a large number of candidate cells. Creative Biogene offers an automated flow cytometry-based screening strategy that delivers efficient screening in a matter of days. We obtain cell count and viability information by using an integrated system of instruments, software and reagents for rapid multiplex analysis, and this combination of information helps us streamline our workflow and filter to the optimal pool of cells.
Creative Biogene offers advanced instrumentation and technology for efficient screening of cell pools. Our experienced team can provide you with the best screening strategy to facilitate your GMP cell line production process. Please feel free to contact us if you have a need, and our experienced specialists will be happy to assist you.
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