Quality improvements of cell membrane chromatographic column

• The methodology of CMC model was fully investigated to improve the technique with the option to obtain more reliable results.
• This is the first time that Paraformaldehyde (PFA) was used in CMC to enhance the efficiency of columns.
• Two novel different CMC models (HSC-T6/CMC and CMMC-7721/CMC) were established.
• This article offers perspectives to be applied for the screening and identification of active compounds in medicines and drugs.

Cell Membrane Chromatography (CMC) is a biological affinity chromatographic method using a silica stationary phase covered with specific cell membrane. However, its short life span and poor quality control was highlighted in a lot of research articles. In this study, special attention has been paid to the disruption, cell load and packing procedure in order to improve the quality of the CMC columns. Hereto, two newly established CMC models, HSC-T6/CMC and SMMC-7721/CMC have been developed and used in this research project. The optimization of the abovementioned parameters resulted in a better reproducibility of the retention time of the compound GFT (RSD < 10%) and improved significantly the quality of the CMC columns. 3.5 × 107 cells were the optimal cell load for the preparation of the CMC columns, the disruption condition was optimized to 5 cycles (400 W and 20 s interval per cycle) by an ultrasonic processor reducing the total time of cell disruption to 1.5 min and the packing flow rate was optimized by applying a linear gradient program. Additionally, 4% paraformaldehyde (PFA) was employed to improve the column quality and prolong the column life span. The results showed that the retention time was longer with PFA treated columns than the ones obtained with the control groups