Protein and Peptide Purification
Using fast protein liquid chromatography (FPLC), the Proteomics Core offers research-scale chromatographic fractionation of proteins from biological fluids, conditioned media, cell lysates and more. Purification of monoclonal antibodies from ascites or hybridoma media, and immunodepletion of abundant proteins from biological fluids, also is available.
Protein separation from complex mixtures is useful and necessary for a variety of research applications, including assay development, receptor binding analysis, biomarker discovery, structural measurements (such as X-ray crystallography), molecular biology studies, and protein and protein complex molecular mass estimation.
The Proteomics Core also provides analytical and semipreparative scale peptide fractionation, purification and analysis using high-pressure liquid chromatography.
All first-time protein and peptide purification requests begin with a consultation meeting to discuss the client's goals, determine purification strategies and set realistic expectations.
Protein purification. A variety of FPLC separation chemistries are available within the Proteomics Core: ion-exchange chromatography, affinity chromatography (including Protein A and Protein G, and Agilent Technologies' Multiple Affinity Removal System for depletion of six or 14 highly abundant proteins in human fluids), size-exclusion chromatography and hydrophobic interaction chromatography.
Also available are other specialty columns, such as hydroxyapatite, as well as various column dimensions for diverse sample amounts and volumes.
- Peptide purification and analysis. Chemistries available in the core are reversed-phase C4, C8 or C18, and hydrophobic interaction liquid chromatography for polar analytes. Dual-wavelength absorbance detectors and fluorescence detection also are available.
The cost for purification or analysis is based on several factors, including the volume to be purified, the number of steps needed for the purification, the chromatographic media expense and the number of hours of technician time needed.
For more information or to initiate a service or project request, email the Proteomics Core.