Quantitative affinity chromatography

There are a number of other chromatographic methods that are closely related to traditional affinity chromatography. For instance, affinity chromatography can be adapted as a tool for studying solute-ligand interactions. This application is known as analytical affinity chromatography, quantitative affinity chromatography, or biointeraction... 

Quality tests, of phenol, 78 753 Quantitative affinity chromatography, 6 404 05... 

Affinity chromatography and related techniques (e.g., thiol chromatography and IMAC) are widely used for preparative isolation because they enable a single protein or class of proteins to be selectively purified from very complex mixtures. They may be occasionally used as analytical tools. For example, protein A affinity chromatography has been used for quantitative analysis of immunoglobulins in ascites fluid.45 Information about surface-accessible histidine and phosphate groups may be obtained using IMAC. 

The first step in biopharmaceutical development is the selection of a clone in a specific cell line. Whole-mass analysis, if possible, is a fairly simple and powerful tool at this stage to verify the successful expression and translation of the desired protein. VanAdrichem et al.65 described the use of MALDI MS to monitor protein expression in several mammalian cell lines like CHO DXB11, CHO SSF3, and hybridomas. Quantitative MALDI-TOF MS measurements of an IgG antibody and insulin during large-scale production in hybridoma cells were comparable to affinity chromatography results. 

A third area of development in carbohydrate l.c. analyses is in the combined techniques (see Section IV,3) and other methods that provide qualitative, as well as quantitative, information about sample constituents, such as high-performance liquid affinity chromatography. The use of specific lectin- and monoclonal antibody-based, stationary phases for analytical and preparative applications is now being considered. The basic concepts of these techniques have been reviewed - and their applications to carbohydrates have been discussed. 

A Lundqvist, P Lundahl. Advantages of quantitative affinity chromatography... 

Winsor, D.J., Quantitative affinity chromatography Recent theoretical developments, in Handbook of Affinity Chromatography, 2nd edn., Hage, D.S., Ed., CRC Press, Boca Raton, FL, 2005, Chap. 23. 

Andrews, R, Kitchen, B.J., and Winzor, D., Use of affinity chromatography for the quantitative study of acceptor-ligand interactions The lactose synthetase system, Biochem. J., 135, 897-900, 1973. 

Dunn, B.M. and Chaiken, I.M., Quantitative affinity chromatography. Determination of binding constants by elution with competitive inhibitors, Proc. Natl. Acad. Set U.S.A., 71, 2382-2385, 1974. 

Kasai, K. and Ishii, S., Quantitative analysis of affinity chromatography of trypsin. A new technique for investigation of protein-ligand interaction, J. Biochem., 77, 261-264, 1975. 

Chaiken, I.M., Quantitative uses of affinity chromatography. Anal. Biochem., 97, 1-10, 1979. 

Phosphorylation on serine, threonine, and tyrosine residues is an extremely important modulator of protein function. Phosphorylation can be analyzed by mass spectrometry with enrichment of compounds of interest using immobilized metal affinity chromatography and chemical tagging techniques, detection of phosphopep-tides using mass mapping and precursor ion scans, localization of phosphorylation sites by peptide sequencing, and quantitation of phosphorylation by the introduction of mass tags (McLachlin and Chait 2001). 

B. Nilsson, Extraction and quantitation of cortisol by use of high-performance liquid affinity chromatography , J. Chromatogr. 276 413-417 (1983). 

Affinity chromatography techniques have shown less utility in analytical testing than in preparative separations for a variety of reasons, including cost and the difficulty of validating consistent operation as the column changes over time. Protein A affinity has been commonly used to quantitate the total antibody content of either ascites or cell culture fluids. To provide guidance in the development of a purification process, specific immunoaffinity resins are either available or can be readily prepared to quantitate the levels of unrelated protein contaminants. To rapidly determine what the active species in a mixture is, a monoclonal antibody that... 

L. H. Reyes, J. M. M. Gayon, J. I. G. Alonso, A. Sanz-Medel, Quantitative speciation of selenium in human serum by affinity chromatography coupled to post-column isotope dilution analysis ICP-MS, J. Anal. Atom. Spectrom., 18 (2003), 11-16. 

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