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Title:Secondary anchor targeted cell release system
Author(s):Ansari, Ali
Department / Program:Bioengineering
Degree Granting Institution:University of Illinois at Urbana-Champaign
Subject(s):Surface Functionalization
RAW 264.7
Cell Isolation
Abstract:The lack of diagnostic tools that can probe individual heterogeneities in patient’s cell receptor expression limits advancement in personalized medicine. These individual differences in receptor quantities can give rise to both intrinsic and acquired resistances to therapeutics, which result in reduced treatment efficacy. In diseases like cancer, where therapeutics have many adverse side effects, noting which drugs have reduced efficacy means the difference between remission and death. Current chemical and physical cell separation methodologies may result in disruption of physiological receptor quantities. These changes in receptor quantities and expression may hide changes that give diagnostic information about tumor progression and environment. Thus separation techniques which hide these changes in receptor expression would be sub-optimal diagnostic tools. Here we describe a functionalization process that facilitates gentle cell capture with subsequent cell release via a secondary, surface-anchoring mechanism. The cellular capture system consists of glass functionalized with APTES, d-desthiobiotin and streptavidin, which when coupled with biotinylated antibodies, such as mCD11b and hIgG, are used to capture mouse macrophages (RAW 264.7) and human breast cancer (MCF7-GFP) cell lines, respectively. Cell release is facilitated through the introduction of biotin, allowing for the enrichment of the cells of interest 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. This engineering advancement is a critical step towards achieving cell isolation platforms for personalized medicine.
Issue Date:2016-04-21
Rights Information:Copyright 2016 Ali Ansari
Date Available in IDEALS:2016-07-07
Date Deposited:2016-05

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