Table of Contents
Chromatography Research International
Volume 2013, Article ID 341036, 6 pages
Research Article

Isolation of Low Abundance Proteins and Cells Using Buoyant Glass Microbubble Chromatography

1Fuzbien Technology Institute, 9700 Great Seneca Hwy, Suite 302, Rockville, MD 20850, USA
2Creatv Microtech Inc., 11609 Lake Potomac Drive, Potomac, MD 20854, USA

Received 21 February 2013; Accepted 23 May 2013

Academic Editor: Joselito P. Quirino

Copyright © 2013 Steingrimur Stefansson et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Conventional protein affinity chromatography relies on highly porous resins that have large surface areas. These properties are ideal for fast flow separation of proteins from biological samples with maximum yields, but these properties can also lead to increased nonspecific protein binding. In certain applications where the purity of an isolated protein is more important than the yield, using a glass solid phase could be advantageous as glass is nonporous and hydrophilic and has a low surface area and low nonspecific protein binding. As a proof of principle, we used protein A-conjugated hollow glass microbubbles to isolate fluorescently labeled neurofilament heavy chain spiked into serum and compared them to protein A Sepharose and protein A magnetic beads (Dynabeads) using an anti-neurofilament protein antibody. As expected, a greater volume of glass bubbles was required to match the binding capacity of the magnetic beads and Sepharose resins. On the other hand, nonspecific protein binding to glass bubbles was greatly reduced compared to the other resins. Additionally, since the glass bubbles are buoyant and transparent, they are well suited for isolating cells from biological samples and staining them in situ.