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Anal Chem. 2012 May 1;84(9):4022-8. doi: 10.1021/ac2033408. Epub 2012 Apr 19.

Channel surface patterning of alternating biomimetic protein combinations for enhanced microfluidic tumor cell isolation.

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1
Department of Bioengineering, University of Illinois at Chicago, Chicago, Illinois 60607, USA.

Abstract

Here, we report a new method for multicomponent protein patterning in a microchannel and also a technique for improving immunoaffinity-based circulating tumor cell (CTC) capture by patterning regions of alternating adhesive proteins using the new method. The first of two proteins, antiepithelial cell adhesion molecule (anti-EpCAM), provides the specificity for CTC capture. The second, E-selectin, increases CTC capture under shear. Patterning regions with and without E-selectin allows captured leukocytes, which also bind E-selectin and are unwanted impurities in CTC isolation, to roll a short distance and detach from the capture surface. This reduces leukocyte capture by up to 82%. The patterning is combined with a leukocyte elution step in which a calcium chelating buffer effectively deactivates E-selectin so that leukocytes may be rinsed away 60% more efficiently than with a buffer containing calcium. The alternating patterning of this biomimetic protein combination, used in conjunction with the elution step, reduces capture of leukocytes while maintaining a high tumor cell capture efficiency that is up to 1.9 times higher than the tumor cell capture efficiency of a surface with only anti-EpCAM. The new patterning technique described here does not require mask alignment and can be used to spatially control the immobilization of any two proteins or protein mixtures inside a sealed microfluidic channel.

PMID:
22482510
PMCID:
PMC3343768
DOI:
10.1021/ac2033408
[Indexed for MEDLINE]
Free PMC Article
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