Monte Carlo Method for Calculating the Probability of Monoclonality of Cell Lines

This study describes the generation of monoclonal producer clones using the ClonePix™ FL robotic system. CHO cells producing the enzyme arylsulfatase B (ASB) were used as an example. As a result, 20 leading clones producing the recombinant lysosomal enzyme ASB were selected. The probability of monoclonality calculated using the formula provided by the developers of the ClonePix™ FL system was 99.94%. However, the probability of monoclonality calculated by the Monte Carlo method was 97.79%. The Monte Carlo method provided a more accurate estimate of the probability of monoclonality, allowing us to make adjustments in the process of clone production and optimize cell seeding in a semi-solid medium by taking into account additional parameters. The probability of monoclonality of producer clones is a necessary parameter for drug registration.