Glycoproteins preparation

Figure 5.7. Steady-state fluorescence polarization versus temperature over viscosity ratio for Trp residues of human aj -acid glycoprotein prepared by acetonic precipitation. Data were obtained by thermal variations in the range 7-35" C. Xex = 300 nm. Xem = 330 nm. Protein concentration is equal to 10 pM. The rotational correlation time determined from the Perrin plot is equal to 13 ns at 20°C is in the same range as that (17 ns) expected for the protein at the same temperature, indicates the presence of residual motions. Also, the extrapolated anisotropy (0.264) is equal to that measured at -35 C (0.267). Source Albani, J. R. 1998, Spectrochimica Acta, Part A. 54, 173-183.

In order to understand the origin of the fluorescence of the Trp residues in a protein, we compared the fluorescence parameters of a i-acid glycoprotein prepared in two different ways 1) by a successive combination of ion displacement chromatography, gel filtration and ion exchange chromatography (ai-acid glycoprotein ) and 2) by ammonium sulfate precipitation (ai-acid glycoprotein ). 

Kanamori A, Inoue S, Xulei Z, Zuber C, Roth J, Kitajima K, Ye J, Troy FA 2nd, Inoue Y (1994) Monoclonal antibody specific for a2,8-linked oligo deaminated neuraminic acid (KDN) sequences in glycoproteins. Preparation and characterization of a monoclonal antibody and its application in immunohistochemistry. Histochemistry 101 333-340... 

Standardization and Testing". The final vaccine is tested for safety, potency, and residual chemicals. Safety includes testing for endotoxin and stetihty. Potency is evaluated by quantitative determination of the amount of hemagglutinin in the vaccine. Antibody to this glycoprotein is associated with protection. The single radial immunodiffusion (SKID) technique is used to standardi2e the mass of this protein in comparison to a reference preparation. 

Proteins. A chiral stationary phase with immobilized a -acid glycoprotein on silica beads was introduced by Hermansson in 1983 [18, 19]. Several other proteins such as chicken egg albumin (ovalbumin), human serum albumin, and cellohy-drolase were also used later for the preparation of commercial CSPs. Their selectivity is believed to occur as a result of excess of dispersive forces acting on the more retained enantiomer [17]. These separation media often exhibit only modest loading capacity. 

There is a wide variety of commercially available chiral stationary phases and mobile phase additives.32 34 Preparative scale separations have been performed on the gram scale.32 Many stationary phases are based on chiral polymers such as cellulose or methacrylate, proteins such as human serum albumin or acid glycoprotein, Pirkle-type phases (often based on amino acids), or cyclodextrins. A typical application of a Pirkle phase column was the use of a N-(3,5-dinitrobenzyl)-a-amino phosphonate to synthesize several functionalized chiral stationary phases to separate enantiomers of... 

A similar plot was prepared using the PSA and reference curve oral absorption data for a series of p-blockers (Fig. 19.7), with predicted oral absorption data plotted against observed bioavailability. The differences in metabolic behavior were clearly apparent, as were the largely unquantified effects of P-glycoprotein (P-gp) and other possible transporters [25]. 

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