Purification of human albumin by the combination of the method of Cohn with liquid chromatography

Tanaka, Kumiko; Shigueoka, E.M.; Sawatani, E.; Dias, G.A.; Arashiro, F.; Campos, T.C.X.B.; Nakao, Hiroshi

doi:10.1590/s0100-879x1998001100003

Cited by 27 publications

(21 citation statements)

References 8 publications

(8 reference statements)

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“…Conventional purification of BSA is based on the original Cohn method [6,7] which involved pH and ethanol precipitations. Recent modifications include anion exchange and size exclusion chromatography [8][9][10][11][12][13][14]. Despite these improvements, BSA preparations are still plagued with contaminants such as DNase I, RNase, proteases, immunoglobulins and endotoxins [23][24][25].…”

Section: Resultsmentioning

confidence: 99%

“…More stringent purification steps are required to make BSA preparations suitable for use in blood transfusion and other clinical applications [4,12]. Even though BSA is in high abundance in cow blood, commercial purification of molecular grade BSA involves several cumbersome and time-consuming steps [14]. One way companies avoid high cost of production is to purify BSA of varying research grades depending on the intended use [23].…”

Section: Resultsmentioning

confidence: 99%

“…Albumin is used as a carrier for several molecules in drug delivery systems, a stabilizing protein to dilute antibodies as well as a blocking agent in various immunoassays and detection protocols [1][2][3][4][5]. Most current approaches to purification of BSA combine the original or modified version of the Cohn method [6,7] with several additional steps such as ion exchange chromatography [8][9][10][11][12][13][14]. The relative inexpensiveness of certain brands of commercial BSA is due to abundance of the protein in bovine serum rather the simplicity, optimization or robustness of the purification strategies.…”

Section: Introductionmentioning

confidence: 99%

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Ammonium sulfate precipitation combined with liquid chromatography is sufficient for purification of bovine serum albumin that is suitable for most routine laboratory applications

Odunuga

Shazhko²

2013

Bio Chem Comp

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The use of bovine serum albumin (BSA) in routine biochemical assays such as restriction enzyme digestion and immunodetection is plagued largely by two common contaminants, DNase and immunoglobulins G (IgGs). Acetylation of BSA to inactivate DNase limits its use as a protein standard due to interference with color development in assays such as Lowry's. In spite of the availability of inexpensive BSA, its purification involves several cumbersome and time-consuming steps. In this work, we employed a modified strategy of ammonium sulfate precipitation coupled with liquid chromatography to purify BSA that is free of DNase and IgGs. Purified BSA tested negative for the presence of DNase and IgGs using DNase and immunodetection assays respectively. We conclude that carefully controlled ammonium sulfate precipitation and liquid chromatography techniques are sufficient to purify BSA suitable for most routine laboratory applications. This purification strategy can yield more than 40 g BSA per liter of serum.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Ammonium sulfate precipitation combined with liquid chromatography is sufficient for purification of bovine serum albumin that is suitable for most routine laboratory applications

Odunuga

Shazhko²

2013

Bio Chem Comp

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“…La albúmina humana se obtiene de plasma o suero humano 9,10 , el método de Cohn ha sido el más utilizado a nivel mundial, en el cual se obtiene la albúmina con una pureza del 95% 9 , en los últimos años este procedimiento ha sido mejorado, alcanzando una pureza de 99.3% 11 . La albúmina se utiliza en pacientes con hipovolémia 12-13 , quemaduras 14 , hemorragias severas, sepsis, cualquier condición de shock con hipoalbuminemia 15 pacientes hemodializados con hipotensión que tienen sobrecarga de líquido y que no pueden tolerar soluciones salinas o grandes volúmenes, síndrome nefrótico, hipoalbuminemia por lesión hepática [16][17] .…”

Section: Introductionunclassified

La Albumina humana su utilización en diferentes patologías y su actual

Parra

2015

BISTUA

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La albumina humana se viene utilizando desde 1945 en diferentes patologías y en los últimos años ha aumentado el número de patologías donde se usa este medicamento, los métodos de purificación a través de todos estos años se han venido mejorando y simplificando, lo que eleva su calidad y permite la reducción de los precios del medicamento. En la Universidad de Pamplona, se ha realizado el proceso completo, desde la simplificación del método de producción, sin pérdida de calidad, como la producción piloto de la albumina para análisis internacional de calidad. Los resultados obtenidos por métodos de alta fidelidad, demuestran una pureza superior al 99.98%, lo que eleva este producto de la Universidad de Pamplona a nivel internacional como un producto con capacidad competitiva en calidad y precio.

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“…All the globulins (alpha, beta and gamma) are removed by the method of Cohn et al (10) and nonspecific hemagglutinin by thermocoagulation (11). Final purification is performed by liquid chromatography using the anion ion-exchange gel DEAE-Sepharose FF (Pharmacia, Uppsala, Sweden) (8,12,13).…”

Section: Introductionmentioning

confidence: 99%

Isolation of bovine plasma albumin by liquid chromatography and its polymerization for use in immunohematology

Tanaka

Sawatani

Shigueoka

et al. 2001

Braz J Med Biol Res

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The aim of the method described here is to remove hemoglobin, the major contaminant in the bovine plasma obtained from slaughterhouses, by adding a mixture of 19% cold ethanol and 0.6% chloroform, followed by fibrinogen and globulin precipitation by the Cohn method and nonspecific hemagglutinin by thermocoagulation. The experimental volume of bovine plasma was 2,000 ml per batch. Final purification was performed by liquid chromatography using the ionexchange gel DEAE-Sepharose FF. The bovine albumin thus obtained presented ³99% purity, a yield of 25.0 ± 1.2 g/l plasma and >71.5% recovery. N-acetyl-DL-tryptophan (0.04 mmol/g protein) and sodium caprylate (0.04 mmol/g protein) were used as stabilizers and the final concentration of albumin was adjusted to 22.0% (w/v), pH 7.2 to 7.3. Viral inactivation was performed by pasteurization for 10 h at 60 o C. The bovine albumin for the hemagglutination tests used in immunohematology was submitted to chemical treatment with 0.06% (w/v) glutaraldehyde and 0.1% (w/v) formaldehyde at 37 o C for 12 h to obtain polymerization. A change in molecular distribution was observed after this treatment, with average contents of 56.0% monomers, 23.6% dimers, 12.2% trimers and 8.2% polymers. The tests performed demonstrated that this polymerized albumin enhances the agglutination of Rho(D)-positive red cells by anti-Rho(D) serum, permitting and improving visualization of the results.

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Purification of human albumin by the combination of the method of Cohn with liquid chromatography

Cited by 27 publications

References 8 publications

Ammonium sulfate precipitation combined with liquid chromatography is sufficient for purification of bovine serum albumin that is suitable for most routine laboratory applications

Ammonium sulfate precipitation combined with liquid chromatography is sufficient for purification of bovine serum albumin that is suitable for most routine laboratory applications

La Albumina humana su utilización en diferentes patologías y su actual

Isolation of bovine plasma albumin by liquid chromatography and its polymerization for use in immunohematology

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