Search results for "SIZE-EXCLUSION CHROMATOGRAPHY"
showing 10 items of 137 documents
Biocompatible polymeric micelles with polysorbate 80 for use in brain targeting.
2008
In this paper, the synthesis and characterization of novel amphiphilic graft copolymers based on an alpha,beta-poly(N-2-hydroxyethyl)-D, L-aspartamide (PHEA) backbone and D, L-polylactic acid (PLA) hydrophobic side chains are reported. These copolymers were obtained starting from PHEA-ethylenediamine (PHEA-EDA), which was functionalized with polysorbate 80 (PS(80)) and/or PLA in order to obtain the PHEA-EDA-PS(80)-PLA and PHEA-EDA-PLA samples, respectively. The degrees of derivatization, DD(PS80) and DD(PLA), of PHEA-EDA-PS80-PLA, calculated by (1)H-NMR, resulted in being 1.2 +/- 0.03 mol% and 0.54 +/- 0.05 mol%, respectively, while that of PHEA-EDA-PLA was found to be 0.60 +/- 0.05 mol%. S…
Separation studies of amino acids, proteins and enzymes on bonded 1,2-dihydroxy-, 1,2-hydroxylamino-and amino silica packings
1979
1,2-dihydroxy-3-propoxypropyl (HPPS), 1-amino-2-hydroxy-3-propoxypropyl (AHPS) and 1-aminoethyl-3-aminopropyl (AEAPS) silica were synthesized by means of both a surface modification procedure (I) and a bulk modification procedure (II). Method (I) gave a surface concentration, α, of functional groups of 2–3 μmole/m2, whereas method (II) gave values up to 5 μmole/m2. Retention times, peak asymmetries and plate heights of thiamine and ascorbic acid eluted with aqueous buffer solutions ranging from pH 5.3 to 9.2 gave only a±5% variation over periods of 12 hours and more. The recoveries of selected enzymes and proteines examined under static and dynamic conditions were between 60% and 100% depen…
Keyhole Limpet Hemocyanin (KLH): Slow In Vitro Reassociation of KLH1 and KLH2 from Immucothel®
1998
Abstract Following our in vitro reassociation of keyhole limpet hemocyanin subunits in the presence of high concentrations (100 mM each) of calcium and magnesium chloride (Harris et al., 1997a, Micron 28, 31–41; 1997b, Micron 28, 43–56), we have now extended our investigations by using a buffer system containing a lower concentration of the two divalent cations (10 mM each). Reassociation of mixed KLH subunits present in the commercially available product Immucothel® was performed using a standardized buffer solution containing 50 mM Tris–HCl, 150 mM NaCl, 10 mM CaCl2 and 10 mM MgCl2 (pH 7.4) over a minimum period of one week, at 4°C. This solution was selected as being close to our KLH sta…
Preparative separation of proteins and enzymes in the mean molecular-weight range of 10,000–100,000 LiChrosorb diol® packing by high-performance size…
1979
Abstract LiChrossorb Diol® packing has been to be well-suited for the separation of proteins and enzymes according to a size-exclusion mechanism in a mean molecular-weight (MW) range between 10,000 and 100,000. Loadability of a small bore column of 6 mm I.D. (A) and a large-bore column of 23.5 mm I.D. (B), both of 250 mm in length, were examined. Defining a 20% decrease of the number of theoretical plates as loadability limit the volume load at constant mass of chymotrypsinogen as representative test solute was ≈ 100 μl for column (A) and 1500 μl for column B at 0.78 · 10-5 g/g of packing for column A and 0.78 · 10-6 g/g of packing for column B, respectively. Mass load to constant injection…
Determination of heavy metal complexes with humic substances by HPLC/ICP-MS coupling using on-line isotope dilution technique
1997
An isotope dilution mass spectrometric (IDMS) method has been developed for the simultaneous determination of the complexes of 11 heavy metals (Ag, Cd, Cu, Mo, Ni, Pb, Tl, U, W, Zn and Zr) with humic substances (HS) by coupling HPLC with ICP-MS and applying the on-line isotope dilution technique. The HPLC separation was carried out with size exclusion chromatography. This HPLC/ICP-IDMS method was applied to samples from a brown water, ground water, sewage and seepage water as well as for a sample containing isolated fulvic acids. The total contents of heavy metals and of their complexes were analyzed in these samples with detection limits in the range of 5–110 ng/L. The analysis of heavy me…
Solution properties of polyelectrolytes. II. Comparison of the effects of both polyion and eluent salt concentrations on the calibrations in aqueous …
1987
The elution behaviour of charged water-soluble polymers (sodium polystyrene sulphonate) in both salt-free and NaCl-containing aqueous mobile phases has been investigated by size-exclusion chromatography. A comparative analysis has been established between the results as a function of polyion and salt concentrations, which demonstrates that the chromatographic behaviour is similar in both cases. The variation of the slopes of the different concentration-dependent calibration curves is discussed in terms of polyelectrolyte conformational changes. Finally, a semi-empirical model has been developed to predict the dependence of the elution volume in either polyion or salt concentration, and its …
Comparative study for separation of aquatic humic-type organic constituents by DAX-8, PVP and DEAE sorbing solids and tangential ultrafiltration: ele…
2005
Aquatic humic-type solutes were separated in parallel from the same fresh water source by four different procedures: non-ionic polymethyl methacrylate (DAX-8) and functional cross-linked polyvinylpyrrolidone (PVP) resins, functional diethylaminoethyl cellulose (DEAE) and tangential ultrafiltration completed with a weakly basic anion exchange resin (IRA-67). The similarity-dissimilarity between the quantities and qualities of the different humic samples is discussed, especially in the light of the original dissolved organic matter (DOM). During the past two decades, a significant progress has occurred in the aquatic humic research due to the so-called hydrophobic-hydrophilic properties posse…
Purification and Characterization of Glutamine Synthetase Isoenzymes from Leaves and Roots of Brassica napus (L.)
1995
Summary The glutamine synthetase enzymes from leaves (GS2) and roots (GSR) of Brassica napus L. have been purified to homogeneity by the application of a three-stage isolation procedure comprising anion-exchange chromatography, adsorption by hydroxylapatite and gel filtration on Sephacryl 5-300 HR. The isoforms of the enzyme show a differential distribution in leaf and root tissues. Elution profiles of hydroxylapatite chromatography showed a distinct behaviour for GS proteins found in leaves and roots. Denaturing SDS-PAGE and Western blot experiments revealed molecular masses of approximately 43.5 and 40.5 kD for GS2 and GSR subunits, respectively. Moreover, kinetic properties determined us…
Calculation of the molecular masses of two newly synthesized thermostable enzymes isolated from thermophilic microorganisms
1995
Two thermostable enzymes synthesized by thermophilic microorganisms were isolated and purified. A thermostable beta-galactosidase was produced in a continuous fermentation process by Bacillus stearothermophilus TP 32 as an intracellular enzyme. After applying different concentration procedures the raw extract enzyme was prepurified on a Sephadex G-200 size exclusion column. The isolated beta-galactosidase fraction was then separated with HPLC on a TSK G 3000 SW size exclusion column to determine the molecular mass based on calibration curves of standard proteins. The other enzyme, a thermostable protease, was synthesized by Bacillus stearothermophilus TP 26 as an extracellular enzyme. After…
Size Exclusion Chromatography of Proteins on Improved Bonded Silica Columns
1985
Publisher Summary This chapter discusses the size exclusion chromatography (SEC) of proteins on improved bonded silica columns. Over the past few years, the use of high performance liquid chromatography (HPLC) has been extended to protein chemistry as a powerful separation and isolation technique. A spectrum of variants providing high resolution potential—ion exchange, reversed phase, hydrophobic interaction, affinity, and size exclusion—is available. Of these, SEC is the simplest with respect to method development and equipment design: proteins are separated according to their size in a short and predictable time span under isocratic conditions. The SEC columns are packed with micro-partic…