Monolithic chromatography media for the large scale separation of human therapeutic cells,
PhD Supervisor(s): Eirini Theodosiou, Christopher Hewitt
Contact Email:C.Milner@lboro.ac.uk
Undergraduate Degree: MEng Chemical Engineering, Loughborough University
PhD Summary
Despite recent advances in regenerative medicine, barriers still exist for large-scale isolation of human therapeutic cells. The ideal purification technique should combine high cell purity, yield and function, with fast processing and affordability. Although magnetic separation methods have been employed extensively, there are several key disadvantages i) high cost ii) particles subject to cell up-take, leading to non-affinity binding, and iii) drawbacks when applied to niche cell populations, such as those requiring multiple tandem separation steps and/or involving combined positive and negative cell selection steps.
The aim of this project is to develop a new generic high-throughput cell affinity selection system based on cast polymer monolithic chromatography media featuring giant convective pores.
In-house-fabricated and commercially acquired (Protista Int. AB, Sweden) polyacrylamide cryogel monoliths were formed by free radical polymerisation. T-lymphocytes (Jurkat line), used for chromatography studies. Investigations include varying polymer and initiator concentrations and adding PEG. Cryogel characterisation by swelling/de-swelling rate and ratio, SEM, ESEM. Additionally, Jurkats were introduced to 1 ml Protista cryogels via a GE Akta chromatography system to analyse the ‘after-flow’ cell viability and chromatographic performance of the cryogels.
Skills and techniques include: Polymer monolith fabrication, GE Akta Prime control and chromatogram analysis, mammalian cell culture, scanning-electron microscopy, environmental scanning-electron microscopy, microtome, flow-cytometry, BET surface analysis.
