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PEDS Advance Access published online on April 12, 2007
Protein Engineering Design and Selection, doi:10.1093/protein/gzm016
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© The Author 2007. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Chimeric protein for selective cell attachment onto cellulosic substrates

Scott J. Craig1, Alan Shu1, Yin Xu2, Frances C. Foong2 and Robert Nordon1,3

1 Graduate School of Biomedical Engineering 2 School of Biotechnology and Biomolecular Sciences, University of New South Wales, Australia

3 To whom correspondence should be addressed. E-mail: r.nordon{at}unsw.edu.au

We have developed a fusion protein (CBD-LG) incorporating a cellulose-binding domain and an antibody binding domain, protein LG, to provide an adaptor molecule for cell separation with regenerated cellulose hollow fiber arrays. A single hollow fiber cell adhesion assay utilizing a CD34+ cell line, KG1a, was used to investigate whether ligand affinity interactions were strong enough for cell attachment and separation. CBD-LG efficiently captured CD34+ cells labeled with the mouse IgG2a monoclonal antibody MHCD3400. However, it was not possible to bind CD34+ cells labeled with an IgG1 antibody (HPCA-2). The low affinity of HPCA-2 for LG was overcome by secondary antibodies: KG1a cells that were dual labeled with HPCA-2 followed by rat anti-mouse IgG1 adhered inside hollow fibers coated with CBD-LG. Alternatively, immobilized rabbit polyclonal anti-mouse IgG1 captured cells labeled with HPCA-2. The development of an adaptor molecule to display recombinant domains at the surface of hollow fibers will be an effective tool to investigate cellular ligand–receptor interactions, a necessary step in the development of hollow fiber bioreactors for manufacture of human cellular products.

Keywords: cell adhesion/cell separation/cellulose binding domain/cellulose hollow fibers/recombinant fusion proteins

Received September 1, 2006; revised January 25, 2007; accepted February 28, 2007.


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