Chromatography boronate affinity

Crude chloroform-methanol-water (30 60 8, v/v) extracts of immunostainedTLC bands were analyzed without further purification by nanoelectrospray low-energy mass spectrometry. The authors showed that this effective PLC/MS-joined procedure offers a wide range of applications for any carbohydrate-binding agents such as bacterial toxins, plant lectins, and others. Phenyl-boronic acid (PBA) immobilized on stationary support phases can be put to similar applications. This technology, named boronate affinity chromatography (BAC), consists of a chemical reaction of 1,2- and 1,3-diols with the bonded-phase PBA to form a stable... 

Typically, the boronic acid group is part of an aminophenyl boronic acid derivative, and this group has been used for bioconjugation and affinity chromatography purposes (Burnett et al., 1980 O Shannessy and Quarles, 1987). A common partner for a phenyl boronic acid group in bioconjugation is the salicylhydroxamic acid (SHA) group (Chapter 17, Section 3) (Reaction 59). 

Boron-aluminum-magnesium alloys, 4 137 Boron analysis, of water, 26 39 Boronate affinity chromatography, 6 403 04... 

Glad, M., Ohlson, S., Hansson, L., Mansson, M.-O., and Mosbach, K., High-performance liquid affinity chromatography of nucleosides, nucleotides and carbohydrates with boronic acid-substituted microparticulate silica, J. Chromatogr., 200, 254-260, 1980. 

Transferrin (from human or bovine serum) [11096-37-0J Mr 80,000. Purified by affinity chromatography on phenyl-boronate agarose followed by DEAE-Sephacel chromatography. The product is free from haemopexin. [Cook et al. AB 149 349 1985 Aisen and Listowsky Annual Reviews of Biochem 49 357 1980). 

Cook, N. D. and Peters, T. J. (1985) Purification of y-glutamyl transferase by phenyl boronate affinity chromatography. Biochim. Biophys. Acta 828,205—212. 

Affinity chromatography Has a wide number of uses and can be applied to the isolation and purification of virtually all biomolecules. Specific applications include nucleic acid purification, protein purification from cell and tissues extracts, and antibody purification from blood serum. There are a number of matrices used for the construct, and some examples of these and their uses are as follows heparin columns to separate cholesterol lipoproteins, lectin columns to separate carbohydrate groups, and phenyl boronate columns to separate glycated haemoglobins. 

Cook N D and Peters T J (1985) Punfication of yglutamyl transferase by phenyl boronate affinity chromatography Biochim Bwphys. ActaSiS, 2.06-11%... 

Boronic acid and its derivatives are another class of nonbiological ligands that have weak binding and are often used under isocratic conditions. These are employed in the method of boronate affinity chromatography, which makes use of the ability of such ligands to form covalent... 

Figure 21. Affinity chromatography of fungal proteins on columns packed with sorbent A) vanillin chemically bonded with the CPG surface B) vanillin chemically bonded with the boron-enriched CPG surface. Peroxidase activity-open rings absorbance at 280 nm-black rings.

Dihydroxyphenylalanine-containing proteins. Boronate affinity chromatography is used in the selective binding of proteins containing 3,4-dihydroxyphenylalanine to a m-phenylboronate agarose column and eluted with IM NH4OAC at pH 10. [Hankus et al. Anal Biochem 150 187 7956.]... 

Subtilisin (from Bacillus subtilis) [9014-01-1] Mr 27,000 (sedimentation equilibrinm) [EC 3.4.21.62], This alkaline protease is purified 211-fold by affinity chromatography using 4-(4-aminophenylazo)phenylarsonic acid complex to activated CH-Sepharose 4B. It is inhibited by 2-phenylethane boronic acid, PMSF, 3,4-dichloroisocoumarin, acetone and benzamide. [Chandraskaren Dhar Anal Biochem 150 141 1985, Schomburg Schomburg Springer Handbook of Enzymes 2nd Edn vol 7 p 286 2002.]... 

Streptozotocin-induced diabetic rats also had high levels of glycated Cu,Zn-SOD in the erythrocytes (K9) and lens, as judged by affinity chromatography on a boronate column (see Fig. 10). As described above, much of the glycated... 

Lyon and Phelps [57] have evaluated various glycan-substituted AH-Sepharose gels to be used for affinity chromatography. Srivastava and Farooqui reported the elution of hyaluronidase from heparin-Sepharose columns using heparin or hyaluronan solutions as eluent [58]. Alternatively, affinity chromatography of hyaluronidase has been performed using concavalin A-Sepharose [58] or Matrex gel phenyl boronate [59] columns. 

Novel tetrahedral boronate complexes have been prepared as potential ligands for affinity chromatography either from the amination (LiN(TMS)2) of a-haloalkylboronates followed by A-acetyl-ation (i.e., (132)) or, alternatively, from the a-iodo derivatives and acetamidine (i.e., (133)) (Scheme 17). The 1,3,2-dioxaborinanes proved to be less stable than the pinanediol derivatives to silica gel. "B NMR as well as other data confirmed their chelated nature (e.g., (132) d 6.8) <92JOM(43i)255>. Spiroborates (e.g. (134)) have also been prepared from boric acid, diols, and enolizable 1,3-dicarbonyl compounds <86JPR755>. Vinylzirconocenes (135) are converted to vinylboronates (136) with chloroboranes <910M3777>. The hydrozirconation of 5-vinylboronates (137) produces the mixed... 

R.R. Maestas, J.R. Prieto, G.D. Kuehn and J.H. Hageman. Polyacryamide-boronate beads saturated with biomolecules A new general support for affinity chromatography of enzymes. 

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