A Method of Targeted Cell Isolation via Glass Surface Functionalization.
One of the limiting factors to the adoption and advancement of personalized medicine is the inability to develop diagnostic tools to probe individual nuances in expression from patient to patient. Current methodologies that try to separate cells to fill this niche result in disruption of physiological expression, making the separation technique useless as a diagnostic tool. In this protocol, we describe the functionalization and optimization of a surface for the cellular capture and release. This functionalized surface integrates biotinylated antibodies with a glass surface functionalized with an aminosilane (APTES), desthiobiotin and streptavidin. Cell release is facilitated through the introduction of biotin, allowing the recollection and purification of cells captured by the surface. This release is done through the targeting of the secondary moiety desthiobiotin, which results in a much more gentle release paradigm. This reduction in harsh reagents and shear forces reduces changes in cellular expression. The functionalized surface captures up to 80% of cells in a single cell mixture and has demonstrated 50% capture in a dual-cell mixture. Applications of this technology to xenografts and cancer separation studies are investigated. Quantification techniques for surface verification such as plate reader and ImageJ analyses are described as well.
Department of Bioengineering, University of Illinois at Urbana-Champaign.
Ali
A
Ansari
Department of Liberal Arts & Sciences, University of Illinois at Urbana-Champaign.
Reema
R
Patel
Department of Bioengineering, University of Illinois at Urbana-Champaign.
Kinsey
K
Schultheis
Department of Biomedical Engineering, Illinois Institute of Technology.
Vesna
V
Naumovski
Department of Bioengineering, University of Illinois at Urbana-Champaign; pii@illinois.edu.
P I
PI
Imoukhuede
D000906:Antibodies; D001710:Biotin; D002469:Cell Separation; D005898:Glass; D064593:Heterografts; D006801:Humans; D007202:Indicators and Reagents; D009369:Neoplasms; D019809:Streptavidin
D000906:Antibodies; D007202:Indicators and Reagents; D001710:Biotin; D019809:Streptavidin
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