Human serum albumin acetate

Fig. 4. Backscattered Raman and ROA spectra of the n-helical protein human serum albumin in H20 (top pair) and the /3-sheet protein jack bean concanavalin A in acetate buffer solution at pH 5.4, together with MOLSCRIPT diagrams (Kraulis, 1991) of their X-ray crystal structures (PDB codes lao6 and 2cna). 

G. E. Means, M. L. Bender, Acetylation of Human Serum Albumin by p-Nitrophenyl Acetate , Biochemistry 1975, 14, 4989-4994. 

Figure 5 Covalent coupling of cyclic peptide moieties to human serum albumin (HSA). The depicted cyclic peptide, C SRNLIDC, in which C denotes the cyclizing cysteine residues, mimics the receptor binding site of PDGF-BB. First, a sulfhydryl group is introduced to the cyclic peptide by a reaction with succinimide-acetyl thioacetate (SATA). The primary amino groups of lysine in HSA are derivitized with maleimide-hexoyl-At-hydroxysuccinimide ester (MHS). Subsequently, the cyclic peptide is coupled to HSA. In this latter reaction, hydroxyl amine is used to remove the protecting acetate group from the sulfhydryl group of the cyclic peptide.

In the presence of human serum albumin, the H spectrum of acetyl-salicyclic acid is specifically shifted and broadened [119]. The interpretation of changes in T, and T2 require several theoretical assumptions. These have been discussed in detail [120] for JV-acetylsulphanilamide and acetate binding to the active site of carbonic anhydrase. It was concluded that the acetyl groups of these inhibitors have a motion additional to that of the enzyme. It can be shown by NMR that acetate binds to two sites on the enzyme, only one of which is inhibitory to esterase activity (methyls are 4.3 and 4.8 A from the metal in the Mn substituted enzyme [121]). Strict care must be taken to avoid paramagnetic impurities when NMR relaxation enhancement by diamagnetic macromolecules is being studied. A preparation of carbonic anhydrase, for example, can contain 0.24 paramagnetic Cu atoms per Zn atom [122]. 

Fig. 2. pH Activity curves of two anodal-moving components separated from concentrated human gastric juice in the Tiselius apparatus. , Major, faster component, mobility 2.99 X 10— cm /volt/sec o, minor, slower component. Electrophoresis was carried out in 33 mM acetate buffer, pH 2.5. Protein concentration, 0.51 g/100 ml. Human serum albumin as substrate temp. 37° time 3 hr. From Taylor (T15). 

I, Fitos, M. Simonyi, and Z. Tegyey, Resolution of warfarin via enhanced stereoselective binding to human serum albumin induced by lorazepam acetate. Chromatography 87 (H. Kalasz and L. S, Ettre, eds.), Akademiai Kiado, Budapest, 1988, p. 205,... 

Irikura, M. Takadate, A. Goya, S. Otagiri, M., 7-alkylaminocoumarin-4-acetic acids as fluorescent-probe for studies of drug-binding sites on human serum-albumin, Chem Pharm Bull 1991, 39, 724-728... 

Final product may be formulated in liquid or freeze-dried form, and virtually all biopharmaceutical products are sterilized by filtration followed by aseptic processing. The most commonly employed excipients include human serum albumin, polysor-bate 20 or 80, mannitol, sucrose or maltose, amino acids (usually glycine, arginine or histidine) and a buffer (often citrate, acetate or phosphate based). 

Fig. 4.5.7. An example of medium pressure rapid ion exchange chromatography of 16 mg of a protein mixture (left-hand figure) and of 18 mg of a mixture containing protein fragments (right-hand chromatogram) on 20x0.8 cm columns of CM-Spheron 20-40 /im (37). Buffers A, 0.01 ammonia and acetic acid (pH 7) B, 0.1 M ammonia + acetic acid (pH 7) C, O.S M buffer of the same composition was 1 M in NaCl D, 2 M NaCI. The sequence of protein peaks corresponds to their isoelectric points pi. Cyanogen bromine fragments of human serum albumin CB4 and CB3 contain 31 and 185 amino acids.

Human serum-albumin consists of three distinct proteids a, P, and 7, coagulating at 73° (163°.4 F.), 77° (170°.6 P.), and 84° (183°.2 P.). The blood of some animals contains but two of these. They are all precipitated, after removal of serum-globulin by saturation with MgSOt, by saturation with KajSOi. Potassium acetate also precipitates them without coagulation. 

FIGURE 22.5 CZE electropherograms of rhEPO alpha formulated with human serum albumin (HSA), (a) without immunodepletion of HSA, (b) after HSA immunodepletion. Separation conditions Uncoated fused-silica capillary 87 cm x 50 p,m i.d. Separation buffer 0.01 M Tricine, 0.01 M NaCl, 0.01 M sodium acetate, 7 M urea, and 3.9 mM putrescine, pH 5.5 pressure injection 30 s at 0.5 psi 25 kV temperature 35 C detection at 214 nm. (Adapted from Lara-Quintanar, P. et al., J. Chromatogr. A, 1153, 227, 2007. With permission.)... 

Figure 11.13 Chromatographic retention behavior of glycoproteins and non-glyco-proteins on the click boronate affinity hybrid monolith. The experimental conditions were as follows phosphate buffer (pH 7.5, 0.1 M) containing 0.3 M NaCl, switched to 0.2 M acetic acid at 12 min, HSA human serum albumin, TF transferring. (Reproduced from ref. 54 with permission. Copyright 2013, Springer.)...

Diethylenetriamine penta-acetic acid (DTPA) may be covalently coupled to proteins to provide hexadentate chelating sites for metal ions. The metal-macromolecule complexes are useful as probes in biological systems, and a variety of metals with useful physical properties can be specifically bound. The potential of the system was demonstrated by the preparation of [ In]DTPA-human serum albumin, which was shown to have an in vivo distribution in mice nearly identical to that of [ I]human serum albumin. 

Kurono, Y., Maki, T., Yotsuyanagi, T., and Ikeda, K. 1979. Esterase-like activity of human-serum albumin— structure-activity-relationships for the reactions with phenyl acetates and para-nitrophenyl esters. Chem Pharm Bull 27 2781-86. 

Means, G.E., Bender, M.L., 1975. Acetylation of human serum albumin by p-nitro-phenyl acetate. Biochemistry 14 (22), 4989-4994. 

Figure 1. Separation of immunoglobulins from human plasma using a mixed-mode ion-exchanger. Column, 7.75 mm x 100 mm, 5 micron BAKERBOND ABx mobile phase, initial buffer (A) 25 mM MES, pH 5.4 final buffer (B) 500 mM (NH4)2S04 plus 20 mM sodium acetate, pH 6.3 gradient. 0% B to 25% B over 60 min flow-rate, 1.0 ml/min detection UV (280 nm), 1.0 AUFS sample, 2 ml (0.5 ml crude human plasma diluted to 2 ml with buffer A. Peaks 1, human serum albumin 2, immunoglobulin fraction (shaded).

Human oxyhemoglobin (oxyHb A) hydrolyzes 4-nitrophenyl acetate at a higher rate than bovine serum albumin [107], It has been proposed that imidazole catalysis by /3-His2 is primarily responsible for the esterase activity, and... 

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