Rapid purification

Biomolecule Separations. Advances in chemical separation techniques such as capillary zone electrophoresis (cze) and sedimentation field flow fractionation (sfff) allow for the isolation of nanogram quantities of amino acids and proteins, as weU as the characterization of large biomolecules (63—68) (see Biopolymers, analytical techniques). The two aforementioned techniques, as weU as chromatography and centrifugation, ate all based upon the differential migration of materials. Trends in the area of separations are toward the manipulation of smaller sample volumes, more rapid purification and analysis of materials, higher resolution of complex mixtures, milder conditions, and higher recovery (69). 

R. J. Booth and J.C. Hodges, Solid-supported reagent strategies for rapid purification of combinatorial synthesis products, Acc Chem Res 32 18-26 1999. 

Rapid purification To dry benzene, alumina, CaH2 or 4A molecular sieves (3% w/v) may be used (dry for 6h). Then benzene is distd, discarding the first 5% of distillate, and stored over molecular sieves (3A, 4A) or Na wire. 

Rapid purification Dry rer/-butanol with CaH2 (5% w/v), distil and store over 3A molecular sieves. 

Rapid purification Distil, discarding the first 10% of distillate or until the distillate is clear. The distilled CCI4 is then stored over 5A molecular sieves. 

Rapid purification Pass through a column of basic alumina (Grade 1, lOg/mL of CHCI3), and either dry by standing over 4A molecular sieves, or alternatively, distil from P2O5 (3% w/v). Use immediately. 

Rapid purification Distil, discarding the forerun. Stand distillate over Grade I alumina (5% w/v) or 4A molecular sieves. 

Rapid purification Reflux over CaH (5% w/v) and distil. Store over 4A molecular sieves. 

Rapid purification Stir over CaH2 (5% w/v) overnight, filter, then distil at 20mmHg. Store the distd DMF over 3A or 4A molecular sieves. For solid phase synthesis, the DMF used must be of high quality and free from amines. 

Rapid purification Check for peroxides (see Chapter 1 and Chapter 2 for test under ethers). Pre-dry with CaCl2 or better over Na wire. Then reflux the pre-dried solvent over Na (1 % w/v) and benzophenone (0.2% w/v) under an inert atmosphere until the blue colour of the benzophenone ketyl radical anion persists. Distil, and store over 4A molecular sieves in the dark. 

Rapid purification Distil, dry over K2CO3, distil again and store over 4A molecular sieves. 

Rapid purification Pass through an alumina column and fractionally distilling, collecting the desired boiling fraction. 

Deschamps, J. R., Miller, C. E., and Ward, K. B. (1995). Rapid purification of recombinant green fluorescent protein using the hydrophobic properties of an HPLC size-exclusion column. Protein Expression and Purification 6 555-558. 

Welch, W.J. Feramisco, J.R. (1985). Rapid purification of mammalian 70,000 Dalton stress proteins Affinity of the proteins for nucleotides. Mol. Cell. Biol. 5, 1229-1237. 

Lyons, R.E., Lesieur, E., Kim, M., Wong, D., Brownlee, A., Pearson, R., and Elvin, C., The development of synthetic genes encoding repetitive resilin-like polypeptides Construct design, bacterial expression and rapid purification. Protein Eng. Design Select., 20(1), 25-32, 2007. 

Sweeney, S.F., Woehrle, G.H. and Hutchison, J.E. (2006) Rapid purification and size separation of gold nanoparticles via diafiltration. Journal of the American Chemical Society, 128, 3190-3197. 

McCartney, J. E., Rapid purification of a recombinant protein using tandem radial flow ion-exchange column chromatography, BioTechniques, 11,648,1991. 

Titani, K., Sasagawa, T., Resing, K., and Walsh, K. A., A simple and rapid purification of commercial trypsin and chymostrypsin by reverse-phase high-performance liquid chromatography, Anal. Biochem., 123, 408, 1982. 

Purify the modified protein from reaction by-products by dialysis or gel filtration using 50 mM sodium phosphate, 0.15 M NaCl, 10 mM EDTA, pH 7.2. Alternatively, centrifugal spin columns containing a desalting resin may be used for rapid purification (Thermo Fisher). 

Zhang, A., Gonzalez, S.M., Cantor, E.J., and Chong, S. (2001) Construction of a mini-intein fusion system to allow both direct monitoring of soluble protein expression and rapid purification of target proteins. Gene 275, 241-252. 

Wenk, S. O. and J. Kruip (2000) Novel, rapid purification of the membrane protein photosystem I by high-performance liquid chromatography on porous materials. J.Chromatogr. BBiomed SciAppl., 737 131-142... 

Rapid Purification Techniques for Drugs and Metabolites in Biological Matrices.3... 

RAPID PURIFICATION TECHNIQUES FOR DRUGS AND METABOLITES IN BIOLOGICAL MATRICES... 

R. J. Booth, J. C. Hodges, Solid-Supported Reagent Strategies for Rapid Purification of Combinatorial Synthesis Products , Acc. Chem Res. 1999,32,18. 

Scheme 2.29 Rapid purification using tagged reagents for organic synthesis.

Tasaka, K Kobayashi, M Tanaka, T and Inagaki, C. (1984) Rapid purification of monoclonal antibody in ascites by high performance ion exchange column chromatography for diminishing non-specific staining. Acta Histochem. Cytochem. 17, 283-286. 

Stammers, D. K., Tisdale, M., Court, S., Parmar, V., Bradley, C., and Ross, C. K. (1991). Rapid purification and char-aderisation of HIV-1 reverse transcriptase and RNase H engineered to incorporate a C-terminal tripeptide alpha-tubihn epitope, f EBBS Lett. 283,298-302. 

Lawson, R. Cook, K.G. Yeaman, S.J. Rapid purification of bovine kidney branched-chain 2-oxoacid dehydrogenase complex containing endogenous kinase activity. FEBS Lett., 157, 54-58 (1982)... 

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