Affinity chromatography media

In this chapter, we will survey the kinds of solid supports (substrates) and surface chemistries currently used in the creation of nucleic acid and protein microarrays. Which are the best supports and methods of attachment for nucleic acids or proteins Does it make sense to use the same attachment chemistry or substrate format for these biomolecules In order to begin to understand these kinds of questions, it is important to briefly review how such biomolecules were attached in the past to other solid supports such as affinity chromatography media, membranes, and enzyme-linked immxm-osorbent assay (ELISA) microtiter plates. However, the microarray substrate does not share certain unique properties and metrics with its predecessors. Principal among these are printing, spot morphology, and image analysis they are the subjects of subsequent chapters. 

Sigma Affinity Chromatography Media. Sigma Chemical Company, St. Louis, Missouri, 1989. 

Derivatives of cellulose continue to be used as supports for new affinity chromatography media and for immobilization of enzymes. Trypsin has been immobilized on 4-aminobenzyl-cellulose. A penicillinase from Bacillus cereus has been immobilized by glutaraldehyde-mediated reaction with aminoethyl-cellulose. Tannin has been activated with cyanogen bromide and coupled with... 

Considerable effort continues to be devoted to the development of new methods of enzyme immobilization and of novel affinity chromatography media. Immunoadsorption has been applied to the isolation of numerous biologically important molecules. Oligosaccharides co-polymerized with acrylamide have been exploited with much success for the isolation of many lectins from different sources. Active immobilized cells have been prepared and their potential applications for industrial processes and in medicine have been demonstrated. 

Specific Affinity chromatography Medium-low Low Very high High... 

Roy, L, Jain, S., Teotia, S. and Gupta, M.N., Evaluation of microbeads of calcium alginate as a fluidized bed medium for affinity chromatography of Aspergillus niger pectinase. Biotech. Prog., 20 (2004) 1490-1495. 

Characteristics desired of a matrix for affinity chromatography resemble in many ways those sought for a molecular sieve medium. They are as follows ... 

In principle any binding protein or receptor site or membrane fragment containing a receptor site can be purified by the use of an affinity medium containing the moiety with which the protein or receptor interacts. The extensive bibliography rapidly building up in this area of application indicates that, as with most other applications in affinity chromatography, the principle appears to be valid. 

Specific binding to antibodies coenzymes, or hormone receptors. The unique property of a protein to recognize and bind to complementary antibodies with high specificity is the basis for immunochemical assays (see Chapter 9). Proteins can also be separated by affinity chromatography, in which a ligand attached to a solid medium provides high selectivity (see Chapter 6). 

To determine if differential glycosylation of fibronectin (Fn) in inflammatory synovial fluid (SF) included expression of an oncofetal epitope (Onf Fn) previously detected only on Fn derived from embryonal or neoplastic tissue [61], Fn was purified from plasma, SF and synoviocyte conditioned medium by affinity chromatography and analyzed by sodium dodeeyl sulfate-polyacrylamide gel electrophoresis and Western blotting using a monoclonal antibody (FDC-6) specific for the Onf Fn. The Onf Fn was not expressed on... 

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