New process accelerates peptide mapping
A liquid chromatography (LC) process developed by Agilent Technologies Inc. (www.agilent.com/chem) separates and identifies protein fragments up to 20 times faster than current methods, reducing the cost and time required for high-resolution peptide mapping of antibodies.
ZORBAX Poroshell 300SB columns are designed to speed peptide mapping of antibodies up to 20 times faster than current methods.
Analysis from peptide mapping—a technique for analyzing proteins by separating and detecting the mixture of fragments generated when a protein is broken up with chemicals or enzymes—can provide information on the original protein's sequence and detect subtle differences between proteins when coupled with mass spectrometry.
While most peptide mapping methods rely on traditional LC columns containing totally porous particles, the Agilent process uses an LC column packed with its proprietary Poroshell—a superficially porous chromatographic media designed to significantly reduce the time required for peptide mapping of human monoclonal antibodies.
The Poroshell packing technology uses particles consisting of solid silica core, surrounded by a thin outer layer of porous silica. The design, Agilent says, allows for higher throughput and flow rates while maintaining sharp peak shape and resolution. During the separation process, the company says proteins and peptides rapidly diffuse into and out of the thin porous shell, then elute rapidly as easily resolved, narrow bands.
Agilent says its ZORBAX Poroshell 300SB columns are available in several internal diameters and bonded phases.