Figure 4
Clonal origin predicts similarity in bioprocess relevant features despite overall similarity (a) Experimental outline to evaluate the cellular production performance of clonal cells which originated from different RMCE events. Clones were randomly selected (confluence threshold) and expanded for testing in ambr15 microbioreactors. (b) Hierarchical clustering of individual producer clones by antibody chain expression in bulk transcriptome profiling. Note the distance of cells, which share the same barcode. Violin plot comparing (c) absolute differences in product quality parameters, (d) metabolite concentrations, and (e) cellular features between unique barcodes (relative clones) as compared to barcodes with ≥ 3 occurrences (sibling clones). Dotted line indicates the arithmetic mean. FDR-adjusted statistical significance was calculated by Wilcoxon rank-sum test. (f) Product quality (main peak measured by CE-SDS) and titer after protein A purification of clonal cells. Clonal cells with identical barcodes are color matched. Barcodes which occurred ≥ 3 times (sibling clones) are highlighted (circle). (g) Principal component analysis (PCA) of bulk transcriptome data from 94 randomly selected clonal cell lines. Cells were sampled at day 10 during a 14-day fed batch process in ambr15 bioreactors. Clonal cells with identical barcodes are color matched.
