Affinity-based chromatography

Haginaka J (2008) Monodispersed, molecularly imprinted polymers as affinity-based chromatography media. J Chromatogr B Anal Technol Biomed Life Sci 866(l-2) 3-13... 

J. Porath, IMAC-immobilized metal affinity based chromatography. 

Protein A chromatography is the most often used affinity-based chromatography in the purification of therapeutic mAbs, and Fc-containing fusion proteins. 

A number of affinity-based or chromatography methods have been used to prefractionate protein samples for 2D electrophoresis. For example, proteins of low abundance can be enriched from crude lysates by affinity-... 

There is no doubt that the technique of chromatography (particularly affinity-based) will continue to make large advances as our understanding of the molecular mechanisms governing protein folding and enzyme-substrate interactions in-... 

We have developed an analogous, but more robust system which is not necessarily constraint by the aforementioned limitations. The obvious extension has been to couple an affinity-based separation with mass spectrometry. Hutchens et al. have shown that affinity probe surfaces can be ust to capture specific protein ligands allowing detection by laser desorption mass spectrometry (. The limitations to their technique have been that the surface area for ligand capture is quite small and salt (or detergent) contaminants are still problematic. Perfusive affinity resins, on the other hand, provide a tremendous surface area for binding. The nature and composition of the solvents required for affinity chromatography, however, are not directly compatible with mass spectrometric analysis. 

Molecular imprinting technologies have found a niche in separation science. MIPs offer a good alternative to proteinaceous receptors and have been successfully used in affinity-based separations. Furthermore, the polymeric nature of the MIPs bears attractive features for applications that could not be achieved by antibodies or enzymes gas chromatography, solid-phase microextraction, etc. As further fundamental research and apphcation efforts are invested in MIP technologies, it is expected that MIPs will soon be commercialized. MIP-SPE is expected to the first commercial application area. 

Many of the techniques used to enrich PTMs at the protein level are applicable at the peptide level. A popular method for enrichment of phosphopeptides is to use an immobilized metal affinity column. The molecular basis for the enrichment is the phosphate affinity to transition metal ions, such as copper, nickel, cobalt, iron, aluminum, gallium, and zinc (142). A comprehensive source for phosphopeptide enrichment strategies can be found in a recent review (143). Lectin-based chromatography is used to enrich for glycopep-tides (144). Lectins are proteins that have an affinity for carbohydrates. With multiple available enrichment options, choosing the right one for the experiment is important. For determining which proteins have a certain PTM, protein-level enrichment can be performed. However, if a type of PTM is of... 

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