Effective Separation for New Therapeutic Modalities Utilizing Temperature-responsive Chromatography

Abstract

In recent years, drug discovery and therapeutics trends have shifted from a focus on small-molecule compounds to biopharmaceuticals, genes, cell therapy, and regenerative medicine. Therefore, new approaches and technologies must be developed to respond to these changes in medical care. To achieve this, we applied a temperature-responsive separation system to purify a variety of proteins and cells. We developed a temperatureresponsive chromatography technique based on a poly(N-isopropylacrylamide) (PNIPAAm)-grafted stationary phase. This method may be applied to various types of protein and cell separation applications by optimizing the properties of the modified polymers used in this system. Therefore, the developed temperature-responsive HPLC columns and temperature-responsive solid phase extraction (TR-SPE) columns can be an effective separation tool for new therapeutic modalities such as monoclonal antibodies, nucleic acid drugs, and cells.

Recent Advances in Poly(N‐isopropylacrylamide) Hydrogels and Derivatives as Promising Materials for Biomedical and Engineering Emerging Applications

“…[119][120][121] Temperature-responsive HPLC columns and temperatureresponsive solid-phase extraction (TR-SPE) columns were found to work as effective separation tools for monoclonal antibodies, nucleic acid drugs, and cells. In another work, [122] a new device for bioanalysis was manufactured using a poly(NIPAAm-co-BMA-co-DMAPAAm) (where BMA is n-butyl methacrylate and DMAPAAm is N,N-dimethylaminopropyl acrylamide) hydrogel modified TR-SPE system, in which silica beads with average diameters ranging from 100 to 150 μm were employed for optimizing the column towards the size of blood cells. Temperaturedependent cell elution behavior was observed in the separation of HL-60 and Jurkat cells (leukemia cells).…”

“…Reproduced with permission. [ 122 ] Copyright 2021, Springer Nature. b) PNIPAAm brush‐modified substrates (PIPAAm brush, as called in the reference) with a terminal cationic group prepared via RAFT polymerization.…”

“…Concept of interaction between polymer-modified surface and cells, protocol of cell separation using TR-SPE column, and corresponding elution profiles of HL-60 and Jurkat cells. Reproduced with permission [122]. Copyright 2021, Springer Nature.…”

Harvesting methods of umbilical cord-derived mesenchymal stem cells from culture modulate cell properties and functions

“…PNIPAAm exhibits reversible hydration/dehydration changes in response to cell culture temperatures near 37 °C [ 30 ]. The unique properties of PNIPAAm have been exploited in the context of biomedical applications such as drug delivery systems [ [31] , [32] , [33] , [34] , [35] , [36] , [37] ], bioanalysis and biosensor devices [ [38] , [39] , [40] , [41] , [42] , [43] , [44] ], nano-actuators [ [45] , [46] , [47] , [48] ], bioseparation tools [ [49] , [50] , [51] , [52] , [53] , [54] ], cell-separation materials [ [55] , [56] , [57] , [58] , [59] ], cell culture substrates [ [60] , [61] , [62] , [63] , [64] , [65] , [66] , [67] , [68] , [69] ], and cell sheet therapy in diverse types of regenerative medicine [ [16] , [17] , [18] , [19] , [20] , [21] , [22] , [24] , [25] , [26] , [27] , [28] ]. For cell sheet preparation, the cells were seeded onto PNIPAAm-modified dishes.…”

Chromatography columns packed with thermoresponsive-cationic-polymer-modified beads for therapeutic drug monitoring

“…Temperature-responsive chromatography using thermoresponsive polymers, such as poly( N -isopropylacrylamide) (PNIPAAm), can overcome this shortcoming because the chromatography system does not use organic solvents in the mobile phase 6 – 8 .…”