Human serum albumin chromatography

Hanai, T., Miyazaki, R., and Kinoshita, T., Quantitative analysis of human serum albumin-drug interactions using reversed phase and ion-exchange liquid chromatography, Anal. Chim. Acta, 378, 77, 1999. 

The ability of proteins to form enantioselective interactions with a large variety of drugs is used in chiral affinity chromatography. Protein CSPs that are most frequently used for the enantioseparation of pharmaceuticals include bovine serum albumin (BSA), human serum albumin... 

DK Lloyd, A Ahmed, F Pastore. A quantitative relationship between capacity factor and selector concentration in capillary electrophoresis and high-performance liquid chromatography evidence from the enantioselective resolution of benzoin using human serum albumin as chiral selector. Electrophoresis 18 958-964, 1997. 

Damsten, M.C., Commandeur, J.N.M., Fidder, A., Hulst, A.G.,Touw, D., Noort, D. and Vermeulen, N.P.E. (2007) Liquid chromatography/tandem mass spectrometry detection of covalent binding of acetaminophen to human serum albumin. Drug Metabolism and Disposition The Biological Fate of Chemicals, 35 (8), 1408-1417. 

Singh, S.S. and Mehta, J. (2006) Measurement of drug-protein binding by immobilized human serum albumin-HPLC and comparison with ultrafiltration. Journal of Chromatography B, 834, 108-116. 

Tiller, P. R. Mutton, I. M. Lane, S. J. Bevan, C. D. 1995. Immobilized human serum albumin liquid chromatography/mass spectrometry as a method of determining drug-protein binding. Rapid Commun. Mass Spectrom., 9,261-263. 

T. A. G. Noctor and I. W. Wainer, The use of displacement chromatography to alter retention and enantioselectivity on a human serum albumin-based HPLC chiral stationary phase A mini review, J. Liquid Chromatogr., 16 183 (1993). 

Pinkerton TC, Koeplinger KA (1990) Determination of warfarin-human serum albumin protein binding parameters by an improved Hummel-Dreyer high performance liquid chromatography method using internal surface reversed-phase columns. Anal Chem 62 2114-2122. 

Machtejevas E and Maruska A. A new approach to human serum albumin chiral stationary phase synthesis and its use in capillary liquid chromatography and capillary electrochromatography. J. Sep. Sci. 2002 25 1303-1309. 

B. Subfile and N. Thuaud, Determination of tryptophan-human serum albumin binding from retention data and separation of tryptophan enantiomers by high-performance liquid chromatography, J. Litj. Chromatogr., 3 299 (1980). 

T. A. G. Noctor, G. F61ix, and I. W. Warner, Stereochemical resolution of enantiomeric 2-aryl propionic acid non-steroidal anti-inflammatory drugs on a human serum albumin based high-performance liquid chromatography chiral stationary phase, Chtmiatogra ia, 31 55 (1991). 

E. Domenici, C. Bertucd, F Salvadori, and I. W. Wainer, Use of a human serum albumin-based chiral stationary phase for high performance liquid chromatography for the investigation of protein binding Detection of the allosteric interaction between warfarin and benzodiazepine binding sites, /. Pharm. Set., 80 164 (1991). 

I. Fitos, M. Simonyi, Z. Tegyey, L. Otvos, J. Kajtar, and M. Kajtar, Resolution by affinity chromatography Stereoselective binding of racemic oxazepam esters to human serum albumin, /. Chmmatogr., 259 494 (1983). 

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,... 

T. A. G. Noctor, C. D. Pham, and 1, W. Wainer, Exploration of the benzodiazepine binding site of human serum albumin by high performance liquid affinity chromatography. Add/. Pharmacol. 

Fig. 1 Examples of affinity chromatography with an epoxy-immobilized polyclonal anti-human serum albumin (HSA) antibody in a 2.1-mm-inner diameter X 30-mm POROS CO column run at 5 mL/min (8000 cm/h) using phosphate-buffered saline for loading and 12 mM HCl with 150 mM NaCl for elution. The sample was 10 jig HSA at 1 mg/mL. (a) shows the first five analyses of a relatively pure sample of HSA, where the first small peak is the unbound contaminant and the larger peak is the elution of the HSA from the affinity column, (b) shows the results of the last 5 analyses from a set of 5000 and (c) shows the calibration curve before (squares) the 5000 analyses and after (triangles).

Immobilized antibodies have also been used extensively in multidimensional liquid chromatography (MDLC) analyses. As shown in Fig. 2, an affinity column with immobilized anti-HSA is used to capture all of the human serum albumin in a sample, allowing all of the other components to flow through to waste. Then, the affinity chromatography column is eluted directly into a size-exclusion column where albumin... 

Kaliszan, R., Noctor, T.A. and Wainer, I.W. (1992b). Quantitative Structure-Enantioselective Retention Relationships for the Chromatography of 1,4 Benzodiazepines on a Human Serum Albumin Based HPLC Chiral Stationary Phase An Approach to the Computational Prediction of Retention and Enantioselectivity. Chromatographia, 33,546-550. 

Interferon AMa-n3. Interferon alfa-n3 is a glycoprotein produced from cultures of human leucocytes treated with Sendai virus. It is purifled initially by chromatography using a mou.se monoclonal antibody that binds to multiple species of human interferon. Subsequent purification involves incubation at 4°C and pH 2 to kill viruses and gel nitration chromatography. It then has a specific activity of about 2 x 108 lU. The drug is supplied in l-mL vials containing I mL of phosphate buffered saline solution, phenol as a preservative. and I mg of human serum albumin as a stabilizer. This solution should be kept at 2 to 8°C. Therapeutic indications and side effects of interferon alfa-n3 are. similar to those described for interferons alfa-2a and alfa-2b. 

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