Purification chromatography

The peptide is removed from the resin by treating the peptide-containing resin with triethylamine in methanol for a longer time. Extensive column chromatography purification, however, is necessary in each case. 

Fig. 21 Chitin binding of 6xHis-tagged resilin with chitin-binding domain (6 H-resChBD) as compared to 6xHis-tagged resilin without chitin-binding domain (6 H-res). T total protein after affinity chromatography purification, B bound protein eluted from chitin beads, UB unbound protein. Reproduced from [187] with permission from The American Chemical Society, copyright 2009...

In place of silica gel, Florisil is also used as the adsorbent in column chromatography. Purification of chlornitrofen using a Florisil column is as follows after installing a column packed with 10 g of Florisil suspended in n-hexane, the sample solution is added continuously to the column and the initial eluate is discarded. A 100-mL volume of diethyl ether-n-hexane (1 19, v/v) is charged to the Horisil column and the eluate is discarded. Chlornitrofen is eluted with 30 mL of this mixture and the eluate is concentrated to dryness before the addition of acetone for GC analysis. ... 

Scheme 33. (I) K[ F]F-K222. MeCN, 110°C, 3 min, then gas chromatography purification (ii) Me2NCH2CH20H, acetone, 100 °C, 10 min, then concentration to dryness and cation exchange Sep-Pak cartridge purification.

Kariluoto, M. S., Vahteristo, L. T, Piironen, V. 1. (2001). Applicability of microbiological assay and affinity chromatography purification followed by high-performance liquid chromatography (HPLC) in studying folate contents in rye. J. Sci. Food Agric., 81, 938-942. 

Milk acid phosphatase has been purified to homogeneity by various forms of chromaotgraphy, including affinity chromatography purification up to 40 000-fold has been claimed. The enzyme shows broad specificity on phosphate esters, including the phosphoseryl residues of casein. It has a molecular mass of about 42 kDa and an isoelectric point of 7.9. Many forms of inorganic phosphate are competitive inhibitors, while fluoride is a powerful non-competitive inhibitor. The enzyme is a glycoprotein and its amino acid composition is known. Milk acid phosphatase shows some similarity to the phosphoprotein phosphatase of spleen but differs from it in a number of characteristics. 

Exercises in conventional column adsorption chromatography. Purification of... 

D Ham C, Ravanat J-L, Cadet J (1998) Gas chromatography - mass spectrometry with high-performance liquid chromatography purification for monitoring the endonuclease Ill-mediated excision of 5-hydroxy-5,6-dihydrothymine and 5,6-dihydrothymine from y-irradiated DNA. J... 

Skorey, K.I., N.A. Johnson, G. Huyer, and MJ. Gresser. 1999. A two-component affinity chromatography purification of Helix pomatia arylsulfatase by tyrosine vanadate. Prot. Expr. Purif. 15 178-187. 

Ketone 16 can be prepared from glucose in 6 steps without extensive chromatography purification (Scheme 10.5).78. V-Aryl-substituted oxazolidinone-containing ketone 18 can be synthesized... 

Veeraraghavan, K., Bernier, A., and Braendli, E. (1991). Sample displacement mode chromatography Purification of proteins by use of a high performance anion exchange column. J. Chromatogr. 541, 207-220. 

Nopper, B., Kohen, F., and Wilchek, M. (1989). A thiophilic adsorbent for the one-step high-performance liquid chromatography purification of monoclonal antibodies. Anal. Biochem. 180, 66-71. 

Knudsen, K. L., Hansen, M. B., Henriksen, L. R., Andersen, B. K., and Lihme, A. (1992). Sulfone-aromatic ligands for thiophilic adsorption chromatography Purification of human and mouse immunoglobulins. Anal. Biochem. 201, 170-177. 

Figure 5 Schematic of a complete multiplexed and integrated instrumental design with eight capillaries. Stars at I, U1, and U2 represent the multiplexed freeze/ thaw valves. The T-assembly is made up of eight pieces of commercial junctions stacked together. These connect to the manifold M1, the SEC (size-exclusion chromatography) purification columns, and the reaction loops. The cross-assembly is made of eight pieces of standard crosses packed together with built-in heaters. V8 is an eight-position motorized titanium valve with a center port. S1 is a two-position motorized PEEK valve. V6 is a six-position motorized PEEK valve. (Reprinted from Ref. 33 with permission.)...

The specific activity of the enzyme was determined in 0.15M lactose solution (0.02M phosphate buffer, pH 7-0). The activity of the enzyme was expressed in terms of units of activity per mg of enzyme. A unit of activity was defined as a p mole of glucose produced per minute. The soluble lactase activity following affinity chromatography purification was 37-1 units/mg. This represented a 4 fold increase in catalytic potency over the specific activity of the crude enzyme preparation (8.9 units/mg). 

FruA is stereoselective and stereocenter 2 is created with the SNuclear Overhauser Effects (NOEs) observed, and the coupling constants measured, the configurations found for compound 11 were IS, 2S, 3R, and 6R. AH the substituents are equatorially positioned on the cyclohexane ring. The (2S,3R)-configurations are consistent with the usually observed diastereoselectivity of FruA In a second exploration [20], the racemic aldehyde 10 was condensed with DHAP after ketal hydrolysis of rac-14 under acidic conditions. In this case, two major isomers 12 and 13 were isolated in 35 and 29% yields after flash chromatography purification. 

Water Preconcentration by ion-exchange chromatography purification by ion-exchange and solvent extraction NAA and U) No data No data Gladney et al. 1983... 

Separation of the target glycosides from the reaction mixture generally requires column chromatography purification. Alternatively, highly fluorinated compounds are readily separated from nonfluorinated compounds by a simple phase separation. Therefore, organic synthesis... 

Figure 18.40 Example of purification by preparative chromatography. Purification of the enantiomers of a benzodiazepinone on 4.6 x 250 mm column of ceUulose tribenzoate coated on 10 Jim silica. Mobile phase, n-hexane-2-propanol (40 60), 0.25 mL/min, at 49oC. (a) Preparative chromatogram, 0.75 mg. (b) Analysis of fractions B (dashed line) and D (soUd line). Reproduced with permission from A. Katti, P. Erlandsson and R. Ddppen, J. Chromatogr., 590 (1992) 127(Figs. 3 and 5).

Figure 2 Cause-and-effect (fish-bone) diagram for affinity chromatography purification.

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