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- Title
Isolation of Low Abundance Proteins and Cells Using Buoyant Glass Microbubble Chromatography.
- Authors
Stefansson, Steingrimur; Adams, Daniel L.; Cha-Mei Tang
- Abstract
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 proteinA-conjugated hollowglassmicrobubbles to isolate fluorescently labeled neurofilament heavy chain spiked into serumand 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.
- Subjects
PROTEIN analysis; CELLS; MICROBUBBLES; CHROMATOGRAPHIC analysis; SEPARATION (Technology); IMMUNOGLOBULINS; BUOYANCY
- Publication
Chromatography Research International, 2013, p1
- ISSN
2090-3502
- Publication type
Article
- DOI
10.1155/2013/341036