Development and characterization of a formulation based covalent conjugation with polyacrylic acid and recombinant major surface antigen (SAG1) of Toxoplasma gondii

2017 ◽  
Vol 42 (6) ◽  
Author(s):  
Rabia Cakir Koc ◽  
Yasemin Budama Kilinc
Keyword(s):  
Vaccine Development ◽  
Surface Antigens ◽  
Water Soluble ◽  
Size Exclusion ◽  
Primary Amines ◽  
Exclusion Chromatography ◽  
Major Surface ◽  
First Time ◽  
Spontaneous Reaction

AbstractIntroduction:Toxoplasmosis is a major disease caused byMethods:PAA and SAG1 were covalently conjugated using a water-soluble carbodiimide crosslinker that activates carboxyl groups for spontaneous reaction with primary amines. To demonstrate conjugation of these molecules in size exclusion chromatography (SEC), fluorescence spectroscopy was used.Results:For the first time, one of the most important surface antigens ofDiscussion and conclusion:The obtained SAG1-PAA conjugate can be used for vaccine development against

scholarly journals Characterization of the oligomeric states of the CK2 α2β2 holoenzyme in solution

2017 ◽  
Vol 474 (14) ◽  
pp. 2405-2416 ◽  
Author(s):  
Graziano Lolli ◽  
Denise Naressi ◽  
Stefania Sarno ◽  
Roberto Battistutta
Keyword(s):  
Active Form ◽  
Mass Spectrometry Data ◽  
Size Exclusion ◽  
Saxs Data ◽  
Deconvolution Analysis ◽  
Exclusion Chromatography ◽  
Kinase Ck2 ◽  
First Time

The regulatory mechanism of protein kinase CK2 has still to be fully clarified. The prevailing hypothesis is that CK2 is controlled by a self-polymerisation mechanism leading to inactive supramolecular assemblies that, when needed, can be disassembled into the α2β2 monomer, the active form of the holoenzyme. In vitro, monomeric α2β2 seems present only at high ionic strengths, typically 0.35–0.50 M NaCl, while at lower salt concentrations oligomers are formed. In the present study, size-exclusion chromatography (SEC), dynamic light scattering (DLS), small-angle X-ray scattering (SAXS) and mutagenesis have been employed for the characterization of the oligomeric states of CK2 in solution. SAXS measurements at 0.35 M NaCl show for the first time the shape of the α2β2 active monomer in solution. At 0.25 M salt, despite single average properties indicating an aggregated holoenzyme, deconvolution analysis of SAXS data reveals an equilibrium involving not only circular trimeric and linear oligomeric (3–4 units) forms of α2β2, but also considerable amounts of the monomer. Together SAXS and mutagenesis confirm the presence in solution of the oligomers deduced by crystal structures. The lack of intermediate species such as αβ2, α or β2 indicates that the holoenzyme is a strong complex that does not spontaneously dissociate, challenging what was recently proposed on the basis of mass spectrometry data. A significant novel finding is that a considerable amount of monomer, the active form of CK2, is present also at low salt. The solution properties of CK2 shown in the present study complement the model of regulation by polymerization.

The degradation of Indulin ATR by Streptomyces: chemical characterization of the water soluble acid precipitable products

1989 ◽  
Vol 67 (2) ◽  
pp. 350-355 ◽  
Author(s):  
P. F. Vidal ◽  
J. Bouchard ◽  
E. Chornet ◽  
R. P. Overend ◽  
H. Giroux ◽  
...  
Keyword(s):  
High Performance ◽  
Protein Complexes ◽  
Chemical Characterization ◽  
Water Soluble ◽  
Size Exclusion ◽  
Produce Water ◽  
Lignin Fraction ◽  
Exclusion Chromatography ◽  
Soluble Acid

Two strains of Streptomyces, S. badius and S. virdosporus, when incubated with kraft lignin (Indulin ATR) produce water soluble acid precipitable compounds. We show by a combination of elemental analysis, amino acid analysis, and FTIR that these are complexes of lignin fragments strongly associated with proteins. The latter, which can be separated by solvent extraction, are added by the microorganisms themselves. The lignin fraction, which represents approximately 60% of the complex weight, consists of lower molecular weight, lower methoxyl content Indulin fragments. KeyWords: lignin, Indulin, lignin–protein complexes, Fourier transform infrared spectrophotometry, high performance size exclusion chromatography, Streptomyces.

scholarly journals Melt-Polymerization of Acrylamide Initiated by Nucleophiles: A Route Towards Highly Branched and Amorphous Polyamide 3

2020 ◽  
Author(s):  
David Edinger ◽  
Hansjörg Weber ◽  
Ema Žagar ◽  
David Pahovnik ◽  
Christian Slugovc
Keyword(s):  
Thermal Analyses ◽  
Ionic Species ◽  
Water Soluble ◽  
Size Exclusion ◽  
Melt Polymerization ◽  
Biomedical Systems ◽  
Exclusion Chromatography ◽  
Nmr Techniques ◽  
High Degree

The melt-polymerization of acrylamide initiated by nucleophiles allows for the preparation of polyamide 3 (PA 3) with a branching factor of about 1.5. The high share of branching units imparts a fully amorphous morphology featuring a low glass transition temperature of 67 °C and renders the polymer water soluble. The disclosed method provides an easy, resource-efficient and green access to a polymer interesting for applications in biological and biomedical systems. The obtained PA 3 was characterized by several NMR-techniques, MALDI-TOF mass spectrometry, size-exclusion chromatography, thermal analyses and powder-X-ray diffractometry. Preparation and characterization of a 15N-marked polymer complemented the elucidation of the polymers structure. Mechanistically, the polymerization can be considered as an aza-Michael polymerization of acrylamide involving zwitter-ionic species as the key intermediates being responsible for the high degree of branching. <br>

scholarly journals Melt-Polymerization of Acrylamide Initiated by Nucleophiles: A Route Towards Highly Branched and Amorphous Polyamide 3

2020 ◽  
Author(s):  
David Edinger ◽  
Hansjörg Weber ◽  
Ema Žagar ◽  
David Pahovnik ◽  
Christian Slugovc
Keyword(s):  
Thermal Analyses ◽  
Ionic Species ◽  
Water Soluble ◽  
Size Exclusion ◽  
Melt Polymerization ◽  
Biomedical Systems ◽  
Exclusion Chromatography ◽  
Nmr Techniques ◽  
High Degree

The melt-polymerization of acrylamide initiated by nucleophiles allows for the preparation of polyamide 3 (PA 3) with a branching factor of about 1.5. The high share of branching units imparts a fully amorphous morphology featuring a low glass transition temperature of 67 °C and renders the polymer water soluble. The disclosed method provides an easy, resource-efficient and green access to a polymer interesting for applications in biological and biomedical systems. The obtained PA 3 was characterized by several NMR-techniques, MALDI-TOF mass spectrometry, size-exclusion chromatography, thermal analyses and powder-X-ray diffractometry. Preparation and characterization of a 15N-marked polymer complemented the elucidation of the polymers structure. Mechanistically, the polymerization can be considered as an aza-Michael polymerization of acrylamide involving zwitter-ionic species as the key intermediates being responsible for the high degree of branching. <br>

Structure of a shear-thickening polysaccharide extracted from the New Zealand black tree fern, Cyathea medullaris

2020 ◽  
Author(s):  
M Wee ◽  
M Mastrangelo ◽  
Susan Carnachan ◽  
Ian Sims ◽  
K Goh
Keyword(s):  
New Zealand ◽  
Uronic Acid ◽  
Average Molecular Weight ◽  
Shear Thickening ◽  
Water Soluble ◽  
Size Exclusion ◽  
Resonance Spectroscopy ◽  
Tree Fern ◽  
13C Nuclear Magnetic Resonance ◽  
Exclusion Chromatography

A shear-thickening water-soluble polysaccharide was purified from mucilage extracted from the fronds of the New Zealand black tree fern (Cyathea medullaris or 'mamaku' in Māori) and its structure characterised. Constituent sugar analysis by three complementary methods, combined with linkage analysis (of carboxyl reduced samples) and 1H and 13C nuclear magnetic resonance spectroscopy (NMR) revealed a glucuronomannan comprising a backbone of 4-linked methylesterified glucopyranosyl uronic acid and 2-linked mannopyranosyl residues, branched at O-3 of 45% and at both O-3 and O-4 of 53% of the mannopyranosyl residues with side chains likely comprising terminal xylopyranosyl, terminal galactopyranosyl, non-methylesterified terminal glucopyranosyl uronic acid and 3-linked glucopyranosyl uronic acid residues. The weight-average molecular weight of the purified polysaccharide was ~1.9×106Da as determined by size-exclusion chromatography coupled with multi-angle laser light scattering (SEC-MALLS). The distinctive rheological properties of this polysaccharide are discussed in relation to its structure. © 2014 Elsevier B.V.

Sign in / Sign up

Export Citation Format

Share Document