Meet the Expert: Sagrario, R&D Program Manager
The selection of stable protein producing cell lines is a tedious process in which normally two substantially different approaches are being pursued in the field. One strategy relies on robot-assisted, high-throughput screening of thousands of transfected expression cells, whereas the other hinges on the stringent selection of only a handful of high producing cells. In both cases, cell clones are traditionally selected based on protein yields prior to cell banking. Upon cell revival, the suitability of each cell clone is further assessed by generating research material to confirm biological activity as well as testing behavior of the cells under manufacturing like conditions. In addition to these pivotal data sets, a final step in selection of the final manufacturing cell line typically involves confirmation of clonal cell stability over at least 60 cell passages in the absence of selection pressure.
At Batavia Biosciences, we have developed a high-throughput cell screening capability that allows the generation of protein material for assessment of biological activity and cell behavior under manufacturing like conditions, at small scale, at the same time and in one system. This down-scaled technology named SCOUT®, allows for the selection of top protein producer clones based on all crucial selection criteria including yield, biological activity, glycan structure, cell growth, population doubling time and cell density. The SCOUT® system makes use of small-scale bioreactor tubes (2-20 mL) coupled to a high throughput purification technology using purification plates, spin filters and spin traps. This production platform is fully integrated with Batavia’s analytical capabilities to quickly gain pivotal insight in protein production and cell behavior.
The SCOUT® technology was used in a program aimed to generate a high expressing recombinant erythropoietin (EPO) expressing CHO cell line. It took the team about 5 weeks, starting with 250 cell lines, to select 40 cell lines based on excellent population doubling time (18-20 hours) and high cell density fed-batch growth (1×107 cells per mL). Material was purified in a single step process with 65-70% recovery and delivered approximately 600-800 μg EPO from the 20 mL cell cultures. This material proved more than sufficient to perform characterization studies as charge analyses (50 μg), NP-HPLC (100 μg), AEX-HPLC (100 μg) and MALDI-MS analyses (50 μg). In addition, 50 μg of material was used to determine biological activity using an in vitro assay with UT-7 cells. In a timeframe of about 10 weeks, the team banked high EPO producing cell clones (~700 mg/L). All relevant glycan structures were present in the product, resulting in high bioactivity well inside the EPO marketed product release range. This example clearly shows the power of high-throughput, small scale manufacturing technology to aid cell selection and quickly anchor multivariate manufacturing processes reducing timelines and costs.
As a company dedicated to help bringing biopharmaceuticals to the market at higher speed, with reduced costs, and with a higher success rate, Batavia Biosciences has vast experience in developing stable protein expressing cell lines. With our team of experienced protein experts, we are well equipped to take on any challenge associated with production of protein-based pharmaceuticals.