Profiling protein-surface interactions of multicomponent suspensions via flow cytometry

Kozak, Darby, Chen, Annie and Trau, Matt (2008) Profiling protein-surface interactions of multicomponent suspensions via flow cytometry. Langmuir, 24 4: 1204-1211. doi:10.1021/la701847x

Author Kozak, Darby
Chen, Annie
Trau, Matt
Title Profiling protein-surface interactions of multicomponent suspensions via flow cytometry
Journal name Langmuir   Check publisher's open access policy
ISSN 0743-7463
Publication date 2008-02-19
Year available 2007
Sub-type Article (original research)
DOI 10.1021/la701847x
Volume 24
Issue 4
Start page 1204
End page 1211
Total pages 8
Editor D. G. Whitten
Place of publication Washington, DC, U.S.A.
Publisher American Chemical Society
Language eng
Subject 030603 Colloid and Surface Chemistry
Abstract This study presents the use of flow cytometry as a high-throughput quantifiable technique to study multicomponent adsorption interactions between proteins and surfaces. Flow cytometry offers the advantage of high-throughput analysis of multiple parameters on a very small sampling scale. This enables flow cytometry to distinguish between individual adsorbent particles and adsorbate components within a suspension. As a proof of concept study, the adsorption of three proteinssbovine serum albumin (BSA), bovine immunoglobulin gamma (IgG) and fibrinogensonto five surfacemodified organosilica microsphere surfaces was used as a model multicomponent system for analysis. By uniquely labeling each protein and solid support type with spectrally distinguishable fluorescent dyes, the adsorption process could be “multiplexed” allowing for simultaneous screening of multiple adsorbate (protein) and adsorbent (particle surface) interactions. Protein adsorption experiments quantified by flow cytometry were found to be comparable to single-component adsorption studies by solution depletion. Quantitative distribution of the simultaneous competitive adsorption of BSA and IgG indicated that, at concentrations below surface saturation, both proteins adsorbed onto the surface. However, at concentrations greater than surface saturation, BSA preferentially adsorbed. Multiplexed particle suspensions of optically encoded particles were modified to produce a positively and negatively charged surface, a grafted 3400 MW poly(ethylene glycol) layer, or a physisorbed BSA or IgG layer. It was observed that adsorption was rapid and irreversible on all of the surfaces, and preadsorbed protein layers were the most effective in preventing further protein adsorption.
Keyword Bovine Serum-albumin
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status UQ
Additional Notes Published on Web 12/08/2007

Document type: Journal Article
Sub-type: Article (original research)
Collection: Australian Institute for Bioengineering and Nanotechnology Publications
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Created: Thu, 03 Sep 2009, 10:29:21 EST by Mr Andrew Martlew on behalf of Aust Institute for Bioengineering & Nanotechnology