Analysis of Intact Proteins in Biotechnology.
Year: 2011
Type: article
Abstract: The demand for separation techniques for intact proteins is increasing with the introduction of a new generation of high resolution mass spectrometers which are able to measure the mass of small to medium size proteins very accurately. Liquid chromatography is a valuable tool for separating these proteins prior to the MS analysis. Intact protein chromatography is most commonly used in a top-down approach in proteomics or to determine expression levels during recombinant protein synthesis. The size of the protein molecule results in very low diffusion coefficients and therefore slow mass transfer in and out of the pore system. A sufficiently large pore diameter is required to minimize the effects of restricted pore diffusion. We show examples of the separation of intact proteins on a column packed with 3μm C8 silica with 1000A pore size. The molecular weight of the protein examples reach from ribosomal proteins (<40kDa) over monoclonal antibodies (∼150kDa) to intact membrane proteins derived from mouse liver.
Source: Europe PMC (PubMed Central)
Authors: Dan DiFeo, Hans‐Jürgen Wirth, Andrew A. Gooley, Ronit Freeman
Language: English
Subfield: Spectroscopy
Field: Chemistry
Domain: Physical Sciences
Open Access status: green
OpenAlex ID: W252705322