An alternative method of large scale plasma fractionation for the lsolation of serum albumin

Schneider, W; Lefèvre, Hans; Fiedler, H.; McCarty, Lee J.

doi:10.1007/bf01633966

Cited by 41 publications

(3 citation statements)

References 30 publications

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“…To date, all protein adsorption studies related to the topic of solution-phase conformational stability have been conducted in fully aqueous conditions, while there is growing attention to the structure and function of proteins in nonaqueous conditions in general, including in areas such as enzymology, pharmaceuticals, and protein purification. , For example, ethanol is a protein precipitant that is widely used in plasma fractionation protocols to yield therapeutic proteins from blood , and can also be used as a solvent to lower the viscosity of antibody formulations . These important applications have spurred interest in understanding how ethanol and other organic solvents affect protein adsorption at solid–liquid interfaces.…”

Section: Introductionmentioning

confidence: 99%

Unraveling How Ethanol-Induced Conformational Changes Affect BSA Protein Adsorption onto Silica Surfaces

et al. 2020

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Protein adsorption at solid−liquid interfaces is highly relevant to a wide range of applications such as biosensors, drug delivery, and pharmaceuticals. Understanding how protein conformation in bulk solution impacts adsorption behavior is fundamentally important and could also lead to the development of improved protein-based coatings. To date, relevant studies have been conducted in aqueous solutions, while it remains largely unknown how organic solvents and more specifically solvent-induced conformational changes might influence protein adsorption. Herein, using the quartz crystal microbalance-dissipation (QCM-D) and localized surface plasmon resonance (LSPR) techniques, we systematically investigated the real-time adsorption behavior of bovine serum albumin (BSA) protein onto silica surfaces in different water−ethanol mixtures ranging from 0 to 60% (v/v) ethanol. The results showed that there was greater protein adsorption at higher ethanol fractions in the 10−30% range, while more complex adsorption profiles were observed in the 40−60% range. The combination of QCM-D and LSPR measurements led us to further identify specific cases in water−ethanol mixtures where washing steps caused densification of the adsorbed protein layer as opposed to typical desorption of weakly adsorbed molecules in aqueous conditions. We discuss mechanistic factors that drive these overall adsorption trends by taking into account how ethanol fraction affects BSA conformation in bulk solution. Together, our findings demonstrate that BSA proteins can adsorb onto silica surfaces across a wide range of water−ethanol mixture conditions, while specific adsorption profiles depended on the ethanol fraction in a manner closely linked to solution-phase conformational properties.

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Section: Introductionmentioning

confidence: 99%

Unraveling How Ethanol-Induced Conformational Changes Affect BSA Protein Adsorption onto Silica Surfaces

et al. 2020

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“…To the first concentrate (through a 30‐kDa cut‐off membrane) was added sodium octanoate (2.3 g per 100 g of protein) and EDTA to a final concentration of 2 mM, the pH was adjusted to 6.5. The temperature of the solution was elevated to 70 °C in 10 min and maintained for 1 h [26–28]. The solution was cooled rapidly in 10 min to 35 °C.…”

Section: Resultsmentioning

confidence: 99%

Albumin purification from human placenta

Cabrera-Crespo

Gonçalves

Martins

et al. 2000

Biotech and App Biochem

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Albumin is the human protein used mainly for therapeutic purposes. Besides the traditionally used plasma, blood from placenta is an alternative source for albumin purification. We describe here an industrial process for purification of albumin from human placenta. The proposed albumin-purification process, for 50 kg of placentas, comprises: (i) extraction of haemolysed blood with saline and solid/liquid separation by basket centrifugation; (ii) selective precipitation of haemoglobin by ethanol/chloroform and precipitate removal by filtration in a press filter; (iii) concentration/diafiltration of the filtrate in a 30 kDa cross-flow ultrafiltration (CFUF) membrane; (iv) thermo-coagulation at 70 degrees C with sodium octanoate/EDTA; (v) treatment with activated charcoal at pH 3; (vi) concentration/diafiltration of the filtrate in a 30 kDa CFUF membrane; (vii) anion-exchange chromatography Q-Sepharose; (viii) hydrophobic-interaction chromatography with phenyl-Sepharose; and (ix) conditioning and pasteurization. The process yields an average of 4.5 g of albumin/kg of placenta with a purity of 97.1% and A(403) of 0.05 (1% protein). The final product passes pyrogen and toxicity tests in vivo and it does not contain polymers or aggregates, even after the accelerated stability test, as judged by gel filtration, as required by the Brazilian Pharmacopoeia.

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“…Briefly, cold ethanol fractionation involves reducing the sample temperature to approximately -5 °C followed by the addition of ethanol and adjusting the solution pH and ionic strength in order to isolate BSA protein [27][28][29] . By contrast, heat-shock fractionation involves heating the sample (typically to ~60 °C or slightly higher) in order to isolate BSA protein and is conducted in the presence of a fatty acid stabilizer (caprylic acid) [30][31][32] . These two fractionation processes can be completed alone or sequentially (cold ethanol, then heat-shock).…”

Section: Evaluation Strategymentioning

confidence: 99%

Quantitative assessment of bovine serum albumin proteins for blocking applications

Junren

Ferhan

Jackman³

et al. 2019

Preprint

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Bovine serum albumin (BSA) is one of the most widely used protein reagents in the scientific community, especially for surface passivation ("blocking") applications in various bioassays. Numerous BSA protein options are commercially available, however, there is scarce information about which ones are preferable for blocking applications.Herein, we conducted biophysical and bioassay measurements to quantitatively compare the conformational, adsorption, and blocking properties of BSA protein reagents that were obtained through six purification methods. Depending on the method, there were significant differences in the conformational and adsorption properties of BSA proteins, mainly due to the presence of fatty acid stabilizers. In turn, we discovered that fatty acidfree BSA proteins exhibit superior blocking performance to fatty acid-stabilized BSA proteins in surface-and nanoparticle-based bioassays. We critically discuss mechanistic factors behind these performance variations and our findings offer a practical framework to guide BSA selection for blocking applications.A common element of bioassays is the need to minimize nonspecific binding of biomolecules in order to maximize assay specificity and sensitivity 1, 2 . To meet this objective, a wide range of reagents, ranging from crude materials such as milk proteins to synthetic nanomaterials such as finely tuned polymers, have been developed to coat target surfaces and to minimize nonspecific binding events-a function that is commonly referred to as

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An alternative method of large scale plasma fractionation for the lsolation of serum albumin

Cited by 41 publications

References 30 publications

Unraveling How Ethanol-Induced Conformational Changes Affect BSA Protein Adsorption onto Silica Surfaces

Unraveling How Ethanol-Induced Conformational Changes Affect BSA Protein Adsorption onto Silica Surfaces

Albumin purification from human placenta

Quantitative assessment of bovine serum albumin proteins for blocking applications

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