Skip to main content

Research Repository

Advanced Search

Removal of Human Leukemic Cells from Peripheral Blood Mononuclear Cells by Cell Recognition Chromatography with Size Matched Particle Imprints

Chester, Rosie; Das, Anupam A. K.; Medlock, Jevan; Nees, Dieter; Allsup, David J.; Madden, Leigh A.; Paunov, Vesselin N.


Rosie Chester

Anupam A. K. Das

Jevan Medlock

Dieter Nees

Profile Image

Dr David Allsup
Senior Lecturer in Haematology and Honorary Consultant

Vesselin N. Paunov


We report a cell recognition chromatography approach for blood cancer cell separation from healthy peripheral blood mononuclear cells (PBMCs) based on sizematched functionalized particle imprints. Negative imprints were prepared from layers of 15 μm polymeric microbeads closely matching the size of cultured human leukemic cells (HL60). We replicated these imprints on a large scale with UV curable polyurethane resin using nanoimprinting lithography. The imprints were functionalized with branched polyethylene imine (bPEI) and passivated by Poloxamer 407 to promote a weak attraction toward cells. When a matching cell fits into an imprint cavity, its contact area with the imprint is maximized, which amplifies the attraction and the binding selectivity. We tested these imprints specificity for depleting myeloblasts from a mixture with healthy human PBMCs in a cell recognition chromatography setup hosting the imprint. The mixture of fixed HL60/PBMCs ratio was circulated over the imprint and at each step the selectivity toward HL60 was assessed by flow cytometry. The role of the imprint length, flow rate, channel depth, and the bPEI coating concentration were examined. The results show that HL60 cells, closely matching the imprint cavities, get trapped on the imprint, while the smaller PBMCs are carried away by the drag force of the flow. Lower flow rates, longer imprints, and interim channel depth favor HL60 specific retention. The bPEI concentration higher than 1 wt % on the imprint made it less selective toward the HL60 because of indiscriminate attraction with all cells. Particle imprint based cell recognition chromatography was able to achieve selective myeloblast depletion from initial 11.7% HL60 (88.3% PBMC) to less than 1.3% HL60 for 3 h of circulation. The cell recognition chromatography with size-matched microbead imprints can be employed as an efficient cell separation technique and potentially lead to alternative therapies for myeloblasts removal from peripheral blood of patients with acute myeloid leukemia.


Chester, R., Das, A. A. K., Medlock, J., Nees, D., Allsup, D. J., Madden, L. A., & Paunov, V. N. (2020). Removal of Human Leukemic Cells from Peripheral Blood Mononuclear Cells by Cell Recognition Chromatography with Size Matched Particle Imprints. ACS Applied Bio Materials, 3(2), 789-800.

Journal Article Type Article
Acceptance Date Jan 6, 2020
Online Publication Date Jan 17, 2020
Publication Date Feb 17, 2020
Deposit Date Feb 17, 2020
Publicly Available Date Jan 18, 2021
Journal ACS Applied Bio Materials
Print ISSN 2576-6422
Electronic ISSN 2576-6422
Publisher American Chemical Society
Peer Reviewed Peer Reviewed
Volume 3
Issue 2
Pages 789-800
Keywords particle imprints, acute myeloid leukemia, HL60, PBMC, cell recognition, chromatography, microbeads
Public URL
Publisher URL


You might also like

Downloadable Citations