Analytical Ultracentrifugation and its use in Biotechnology

1994 ◽  
pp. 261-297 ◽  
Author(s):  
Steven J. Shire
Keyword(s):  
Analytical Ultracentrifugation

The Nature of Rapidly Extracted Phycocyanin from the Blue-Green Alga Plectonema Boryanum

1971 ◽  
Vol 29 ◽  
pp. 366-367
Author(s):  
M. Kessel ◽  
R. MacColl
Keyword(s):  
Self Assembly ◽  
Analytical Ultracentrifugation ◽  
Uranyl Acetate ◽  
The Self ◽  
Major Protein ◽  
Subunit Structure ◽  
Blue Green Alga ◽  
Thin Sections ◽  
Blue Green Algae ◽  
Cacodylate Buffer

The major protein of the blue-green algae is the biliprotein, C-phycocyanin (Amax = 620 nm), which is presumed to exist in the cell in the form of distinct aggregates called phycobilisomes. The self-assembly of C-phycocyanin from monomer to hexamer has been extensively studied, but the proposed next step in the assembly of a phycobilisome, the formation of 19s subunits, is completely unknown. We have used electron microscopy and analytical ultracentrifugation in combination with a method for rapid and gentle extraction of phycocyanin to study its subunit structure and assembly.To establish the existence of phycobilisomes, cells of P. boryanum in the log phase of growth, growing at a light intensity of 200 foot candles, were fixed in 2% glutaraldehyde in 0.1M cacodylate buffer, pH 7.0, for 3 hours at 4°C. The cells were post-fixed in 1% OsO4 in the same buffer overnight. Material was stained for 1 hour in uranyl acetate (1%), dehydrated and embedded in araldite and examined in thin sections.

Bovine Platelet Proteins II. Purification of Platelet Fibrinogen

1969 ◽  
Vol 21 (03) ◽  
pp. 419-427 ◽  
Author(s):  
N. O Solum ◽  
S Łopaciuk
Keyword(s):  
Analytical Ultracentrifugation ◽  
Insoluble Fraction ◽  
Disc Electrophoresis ◽  
Plasma Fibrinogen ◽  
Starch Gel Electrophoresis ◽  
Freezing And Thawing ◽  
Insoluble Material ◽  
High Resolution Power ◽  
Starch Gel ◽  
Platelet Proteins

Summary1. Platelet fibrinogen has been purified from washed bovine platelets. The procedure was based on the methods for purification of plasma fibrinogen by fractionated precipitations and extractions with ethanol and glycine below 0°, and precipitation of proteins by dimethylformamide at 0°.2. The platelet extract obtained by freezing and thawing of the cells, freed from insoluble material by centrifugation at 23,000 x g for 30 min, contained 0.22 ±0.003mg fibrinogen per 109 platelets. Total protein of this fraction was 0.77 ±0.08 mg per 109 platelets whereas that of the insoluble fraction was 0.79 ±0.09 mg per 109 platelets.3. The most purified platelet fibrinogen fraction contained 91-98% of the protein in a thrombin-clottable state. The yield was approx. 20%. It showed homogeneity in analytical ultracentrifugation, in immunoelectrophoresis using an antiserum produced by immunization of rabbits against platelet extract, and in starch gel electrophoresis using a discontinuous system of Tris HCl and borate buffers offering a high resolution power towards the platelet proteins. Polyacrylamide disc electrophoresis revealed two to three faint lines behind the main fibrinogen line. At least one such line was also observed with purified plasma fibrinogen.

A Cross-Method Comparison of Sub-10 nm Nanoparticle Size Distribution

2019 ◽  
Author(s):  
Ye Yang ◽  
Suiyang Liao ◽  
Zhi Luo ◽  
Runzhang Qi ◽  
Niamh Mac Fhionnlaoich ◽  
...  
Keyword(s):  
Size Distribution ◽  
Analytical Ultracentrifugation ◽  
Statistical Significance ◽  
Method Comparison ◽  
Size Determination ◽  
Size Dependent ◽  
X Ray Scattering ◽  
Transmission Electron ◽  
Gold Nps ◽  
Reliable Representation

Accurate nanoparticle (NP) size determination is essential across research domains, with many functions in nanoscience and biomedical research being size-dependent. Although transmission electron microscopy (TEM) is capable of resolving a single NP down to the sub-nm scale, the reliable representation of entire populations is plagued by challenges in providing statistical significance, predominantly due to limited sample counts, suboptimal preparation procedures and operator bias during image acquisition and analysis. Meanwhile alternative techniques exist, but reliable implementation requires a detailed understanding of appendant limitations. Herein, conventional TEM is compared to the size determination of sub-10 nm gold NPs in solution by small-angle X-ray scattering and analytical ultracentrifugation. Form-free Monte Carlo fitting of scattering profiles offers access to a direct representation of the core size distribution while ultracentrifugation sedimentation velocity analysis provides information of the hydrodynamic size distribution. We report a comparison of these three methods in determining the size of quasi-monodisperse, polydisperse and bimodal gold nanoparticles of 2 – 7 nm and discuss advantages and limitations of each technique.

Lipoprotein classification by analytical ultracentrifugation

Pathology ◽  
1982 ◽  
Vol 14 (2) ◽  
pp. 165-171
Author(s):  
Roy Bickerstaffe ◽  
F.B. Desmond
Keyword(s):  
Analytical Ultracentrifugation

scholarly journals Terpene polyacrylate TPA5 shows favorable molecular hydrodynamic properties as a potential bioinspired archaeological wood consolidant

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Michelle Cutajar ◽  
Fabrizio Andriulo ◽  
Megan R. Thomsett ◽  
Jonathan C. Moore ◽  
Benoit Couturaud ◽  
...  
Keyword(s):  
Molar Mass ◽  
Analytical Ultracentrifugation ◽  
Gel Filtration ◽  
Axial Ratio ◽  
Sedimentation Equilibrium ◽  
Archaeological Wood ◽  
Molecular Hydrodynamic ◽  
Pine Trees ◽  
Minimal Concentration ◽  
Hydrogen Bonding Potential

AbstractThere is currently a pressing need for the development of novel bioinspired consolidants for waterlogged, archaeological wood. Bioinspired materials possess many advantages, such as biocompatibility and sustainability, which makes them ideal to use in this capacity. Based on this, a polyhydroxylated monomer was synthesised from α-pinene, a sustainable terpene feedstock derived from pine trees, and used to prepare a low molar mass polymer TPA5 through free radical polymerisation. This polymer was extensively characterised by NMR spectroscopy (chemical composition) and molecular hydrodynamics, primarily using analytical ultracentrifugation reinforced by gel filtration chromatography and viscometry, in order to investigate whether it would be suitable for wood consolidation purposes. Sedimentation equilibrium indicated a weight average molar mass Mw of (4.3 ± 0.2) kDa, with minimal concentration dependence. Further analysis with MULTISIG revealed a broad distribution of molar masses and this heterogeneity was further confirmed by sedimentation velocity. Conformation analyses with the Perrin P and viscosity increment ν universal hydrodynamic parameters indicated that the polymer had an elongated shape, with both factors giving consistent results and a consensus axial ratio of ~ 4.5. These collective properties—hydrogen bonding potential enhanced by an elongated shape, together with a small injectable molar mass—suggest this polymer is worthy of further consideration as a potential consolidant.

scholarly journals Dynamic range multiwavelength particle characterization using analytical ultracentrifugation

Nanoscale ◽  
2016 ◽  
Vol 8 (14) ◽  
pp. 7484-7495 ◽  
Author(s):  
Johannes Walter ◽  
Wolfgang Peukert
Keyword(s):  
Dynamic Range ◽  
Analytical Ultracentrifugation ◽  
Particle Characterization

An analytical ultracentrifugation based study on the conformation of lambda carrageenan in aqueous solution

2013 ◽  
Vol 97 (1) ◽  
pp. 203-209 ◽  
Author(s):  
Fahad M. Almutairi ◽  
Gary G. Adams ◽  
Mehmet S. Kök ◽  
Christopher J. Lawson ◽  
Roland Gahler ◽  
...  
Keyword(s):  
Aqueous Solution ◽  
Analytical Ultracentrifugation
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