Search results for "SIZE-EXCLUSION CHROMATOGRAPHY"

Keyhole limpet haemocyanin (KLH): Purification of intact KLH1 through selective dissociation of KLH2

1995

Abstract Keyhole limpet haemocyanin (KLH) from almost all newly captive animals contains a mixture of KLH1 and KLH2. We show that the dissociation of KLH2 can be produced during EM specimen preparation by the negative staining-carbon film (NS-CF) procedure and in solution by ammonium molybdate-PEG solutions at slightly acidic pHs. The KLH2 multidecamers split apart in the pH range 7.5-6.5 and in the pH range 6.5-6.0 the individual decamers break open and start to dissociate. At pH 5.9 the dissociation of KLH2 yields predominantly a mixture of single subunits and what appear to be subunit dimers. Over the pH range 7.0-5.7 the KLH1 didecamer remains stable. Separation of intact KLH1, in the f…

Fluorescence of soil humic acids and their fractions obtained by tandem size exclusion chromatography-polyacrylamide gel electrophoresis

2002

Humic acids (HAs) extracted from soils of different origin (chernozem, ferralsol and ranker) and their fractions (A, B and C+D) obtained by tandem size exclusion chromatography–polyacrylamide gel electrophoresis were investigated by steady-state fluorescence spectroscopy in the emission mode. Independently of HA source, high molecular size fractions A and B are shown to be weakly fluorescent. The main fluorophores, especially those emitting at long wavelength (around 500–510 nm), are contained in the polar and low molecular size fractions C+D. As indicated by the observed pH effect, aromatic structures bearing carboxylate and OH substituents may be involved in these longer wavelength emissi…

Solution properties of polyelectrolytes. III. Effect of sodium polystyrene sulfonate concentration on viscometric and size exclusion chromatographic …

1988

The effect of sodium polystyrene sulfonate concentration on the shape of its molecules in aqueous solution at different NaNO3 ionic strengths has been analyzed by viscometry and size-exclusion chromatography (s.e.c). An equation has been developed which predicts the intrinsic viscosity, [η] p.cp.cs, at finite concentration of both polyion (cp) and electrolyte (cs). The experimental results obtained by both techniques can be accounted for in terms of the theory. Several factors involved in the elution mechanism have been considered and the variation of the slopes of s.e.c. calibration curves with cp and cs has been discussed in terms of polyion conformation changes.

Solution properties of polyelectrolytes. I. Exclusion chromatography of sodium polystyrene sulphonate in salt-free water as eluent

1987

The effect of concentration of sodium polystyrene sulphonate on the elution volume in exclusion chromatography has been investigated using salt-free water as eluent. A non-linear increase in the elution volume has been observed with increasing injected sample concentration. Concentration-dependent calibrations have been obtained which approach the universal calibration for uncharged polymers (dextrans) as polyelectrolyte concentration increases. The separation mechanisms are interpreted in terms of electrostatic and hydrodynamic interactions. A semi-empirical model is proposed to predict concentration effects of linear polyions in exclusion chromatography and good agreement has been found b…

Gel permeation chromatography, calibration, and distributions of cellulose trinitrates

1967

Molar Mass Distribution and Size of Hydroxyethyl Starch Fractions Obtained by Continuous Polymer Fractionation

2002

By the use of continuous polymer fractionation (CPF) the initial polymer can be separated into fractions of different molar masses, which makes it possible to obtain hydroxyethyl starch (HES) fractions tailor-made for specific application. Two samples of HES (HES A and HES B) were fractionated by means of CPF. By size-exclusion chromatography-multi-angle laser light-scattering-differential refractive index (SEC/ MALLS/DRI) measurements it was shown that CPF is able to remove the low-molar-mass components and to adjust the samples to various desired molar masses with lower polydispersities than the original samples. In terms of the weight-average mean molar mass M w , the sol fractions have …

Characterisation of disaccharide fragments from the enzymatic digestion of heparin by liquid secondary ion mass spectrometry

1988

Disaccharide fragments resulting from the enzymatic digestion of heparins with heparinase have been purified by gel filtration chromatography and directly analyzed by positive and negative ion liquid secondary ion mass spectrometry (LSIMS). Following the chromatographic purification from excess sodium salt, the mass spectra of di- and tetrasaccharide fragment mixtures enabled the identification of up to three covalently bound sulfate moieties per glycosaminoglycan-disaccharide unit, by means of their molecular ions, containing the corresponding alkali-counterions.

The binding of intravenous and oral biliary contrast agents to human and bovine serum albumin

1978

The binding of two homologous series of oral and intravenous biliary contrast agents to human and bovine serum albumin was investigated using the gel filtration technique. All intravenous compounds are bound to human serum albumin via one high affinity and several low affinity binding sites. Within the concentration range investigated, about 3--5 high affinity binding sites for the oral compounds were found on human serum albumin. In general, the intravenous compounds have a greater affinity for human serum albumin than the oral compounds. No significant differences were found for the binding of the oral compounds to human or bovine serum albumin, while the intravenous compounds have a high…

Mass spectrometric investigations of the kinetic stability of chromium and copper complexes with humic substances by isotope-labelling experiments

1999

Isotope-labelling exchange experiments were carried out to investigate the kinetic stability of Cr(III) complexes with humic substances (HS). To compare the results with those of an ion, not expected to form kinetically stable HS complexes with respect to its electron configuration, Cu(II) was investigated under the same conditions. HS solutions of different origin were therefore spiked with 53Cr(III) or 65Cu(II) after saturation of HS with chromium and copper of natural isotopic composition. In fractions of metal/HS complexes with different molecular weight, obtained by ultrafiltration and HPLC/ICP-MS using size exclusion chromatography (SEC), respectively, the isotope ratios of chromium a…

Organobase-Catalyzed Synthesis of Multiarm Star Polylactide With Hyperbranched Poly(ethylene glycol) as the Core

2011

Multiarm star copolymers consisting of the polyether-polyol hyperbranched poly(ethylene glycol) (hbPEG) as core and poly(L-lactide) (PLLA) arms are synthesized via the organobase- catalyzed ring-opening polymerization of lactide using hbPEG as a multifunctional macroinitiator. Star copolymers with high molecular weights up to 792 000 g mol−1 are prepared. Detailed 2D NMR analysis provides evidence for the attachment of the PLLA arms to the core and reveals that the adjustment of the monomer/initiator ratio enables control of the arm length. Size exclusion chromatography measurements show narrow molecular weight distributions. Thermal analysis reveals a lower glass transition temperature, me…