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Effective cell separation utilizing poly(N-isopropylacrylamide)-grafted polypropylene membrane containing adsorbed antibody

Paper ID Volume ID Publish Year Pages File Format Full-Text
22415 43276 2008 5 PDF Available
Title
Effective cell separation utilizing poly(N-isopropylacrylamide)-grafted polypropylene membrane containing adsorbed antibody
Abstract

We previously developed a cell separation method using a poly(N-isopropylacrylamide)-grafted polypropylene (PNIPAAm-g-PP) membrane containing an adsorbed monoclonal antibody (mAb). The purpose of this study is to elucidate the cell separation mechanism in detail and to design an optimal method. As the grafting yield of PNIPAAm increased, the level of the adsorption of IgG1 and cell adhesion to the membrane decreased. After BSA was adsorbed to a PNIPAAm-g-PP membrane at 6°C, where PNIPAAm was hydrophilic, a small amount of IgG1 was adsorbed to the membrane at 37°C, where PNIPAAm was hydrophobic. The desorption of the adsorbed IgG1 was not enhanced even though temperature was reduced to 10°C, where PNIPAAm was hydrophilic. These results indicate that the antibody adsorbed to the intact PP surface of the membrane predominantly contributes to the capture of target cells through the antigen-antibody reaction and that a thermoresponsive transition of PNIPAAm contributes to the detachment of the captured cells. The total number of cells recovered from a PNIPAAm-g-PP membrane containing the adsorbed mAb decreased as the grafting yield increased. A PNIPAAm-g-PP membrane with a 1.7% grafting yield containing adsorbed anti-human CD34 mAb enriched CD34-positive KG-1a cells to 85% from a 1:1 cell suspension of KG-1a cells and CD34-negative Jurkat cells.

Keywords
poly(N-isopropylacrylamide); polypropylene membrane; grafting yield; cell separation; antibody
First Page Preview
Effective cell separation utilizing poly(N-isopropylacrylamide)-grafted polypropylene membrane containing adsorbed antibody
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 105, Issue 3, March 2008, Pages 221–225
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering