Single cell cloning workflow for cell line development
An optimized workflow for monoclonal antibody production
Introduction
In the biopharmaceutical industry, many biological drugs are produced in mammalian cells. Cell line development (CLD) is used to determine which cell lines have the highest recombinant protein production and are also stable during large-scale manufacturing.
In the CLD workflow, cell lines undergo incremental scaling of culture from static to shaker flask expansion and further scaling up in bioreactors. Cell culture in early CLD is currently limited to static format as the size of the well plates and number of cells limit the ability to agitate the culture.
Performance data
The C.BIRD microbioreactor improved cell growth at low seeding densities by approximately 2.1 to 2.7 times in 96-well plates compared to static culture. Likewise, cell density was improved by approximately 1.3 to 1.8 times in 24-well plates compared to static culture. Meanwhile, cell viability was not affected.
Improvement of clonal expansion workflow
The workflow of single-cell cloning starts from 96-well plate culture.
(A) Schematic photo of the cell confluency (5%) of selected wells in clonal expansion workflow starts from 96-well plate culture. (B-E) The viable cell number and cell viability of clonal expansion workflow started from 96-well plate. (F) The cell number of individual clone of day 14 and day 18.
Applying the C.BIRD microbioreactor in the single cell cloning workflow starting from a 96-well plate can increase the average viable cell number in the 96-well culture scale by 1.74 times and following 24-well culture scale by 1.65 times compared to traditional static culture.
The workflow of single-cell cloning starts from 384-well plate culture.
(A) Schematic photo of the cell confluency (40%) of selected wells in clonal expansion workflow starts from 384-well plate culture. (B-E) The viable cell number and cell viability of clonal expansion workflow started from 384-well plate. (F) The cell number of individual clone of day 14 and day 18.
Applying the C.BIRD microbioreactor in the SCC workflow starting from a 384-well plate can increase the average viable cell number in the 96-well culture scale by 1.25 times and following 24-well culture scale by 1.84-times compared to traditional static culture.
