Process for isolation of plasma or serum proteins
Inventors
Assignees
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Publication Number
US-9624260-B2
Publication Date
2017-04-18
Expiration Date
Abstract
The present invention provides a process for the isolation of one or more proteins) from a protein solution. The process comprising the steps of: a) providing a protein solution comprising one or more specific proteins) and having a preset pH and a preset ionic strength or conductivity, b) applying the protein solution to a packed bed or expanded bed column comprising an adsorbent, and c) obtaining one or more proteins) from the column; wherein the protein solution has been supplemented with an alcohol.
Core Innovation
The invention provides a process for large-scale isolation of one or more proteins from a protein solution obtained from human blood and/or a solution derived from human blood, where the protein solution has not been supplemented with an alcohol. The process includes adjusting the pH and adjusting the ionic strength or conductivity of the protein solution, followed by adsorption on an adsorption column comprising a specific adsorbent particle or a functionalised matrix polymer.
One or more proteins are bound to the adsorbent and are separated from a non-bound material fraction containing non-bound protein(s). Bound proteins are obtained by subjecting the adsorbent to at least one elution buffer, and the resulting bound material fraction is obtained by elution of proteins bound to the adsorbent.
The adsorbent comprises a particle with at least one high density non-porous core surrounded by porous material, with a particle density of at least 1.5 g/ml and a mean volume particle diameter of at most 150 μm, or a functionalised matrix polymer carrying a plurality of covalently attached functional groups comprising an aromatic or heteroaromatic ring-system and one or more acidic groups.
Claims Coverage
The independent claims cover a large-scale protein isolation workflow from human blood-derived protein solutions without alcohol supplementation. The inventive features center on controlled pH and ionic strength or conductivity, specific adsorbent chemistry or particle structure, and fractionation into bound and non-bound material fractions with elution and further processing.
Alcohol-free human blood-derived protein solution with pH and ionic strength or conductivity adjustment
Adjusting the pH of the solution to a pH in the range of 3 to 10; adjusting the ionic strength or conductivity of the protein solution to an ionic strength in the range of 0.0001 to 12 or a conductivity in the range of 0.01 to 1000 mS/cm; the protein solution has not been supplemented with an alcohol.
Adsorbent particle with high density non-porous core and porous material
Applying the protein solution to an adsorption column comprising an adsorbent that comprises a particle with at least one high density non-porous core surrounded by a porous material, with particle density of at least 1.5 g/ml and mean volume particle diameter of at most 150 μm, whereby one or more proteins are bound to the adsorbent; obtaining a non-bound material fraction; and subjecting the adsorbent to at least one elution buffer to elute at least one of the proteins bound to the adsorbent.
Functionalised matrix polymer with covalently attached aromatic or heteroaromatic and acidic groups
Applying the protein solution to an adsorption column comprising an adsorbent that comprises a functionalised matrix polymer carrying a plurality of covalently attached functional groups comprising an aromatic or heteroaromatic ring-system and one or more acidic groups, whereby one or more proteins are bound to the adsorbent; obtaining a non-bound material fraction; and subjecting the adsorbent to at least one elution buffer to elute at least one of the proteins bound to the adsorbent.
Sequential elution with one or more additional elution buffers
Providing a protein solution having a preset pH and a preset ionic strength or conductivity; contacting the protein solution with an adsorbent comprising a particle with at least one high density non-porous core surrounded by a porous material, whereby one or more proteins is bound to the adsorbent; subjecting the adsorbent to an elution buffer to elute at least one of the proteins bound to the adsorbent; and subjecting the adsorbent to one or more additional elution buffers to elute one or more remaining proteins bound to the adsorbent.
Across the independent claims, the invention is directed to large-scale protein isolation from human blood-derived protein solutions without alcohol supplementation, using adsorption under controlled pH and ionic strength or conductivity onto either a high density non-porous core/porous material particle or a functionalised matrix polymer carrying aromatic or heteroaromatic and acidic groups, followed by separation into bound and non-bound fractions, elution of bound proteins, and further processing of the non-bound fraction to obtain non-bound protein(s).
Stated Advantages
Large-scale isolation of one or more proteins from human blood or a solution derived from human blood without supplementing the protein solution with an alcohol.
Documented Applications
Isolation of proteins such as albumin, IgG, fibrinogen, α-1-proteinase inhibitor (α-1-PI), α-1-acid glycoprotein (orosomucoid), and coagulation and anti-coagulation factors including Factor II, Factor V, Factor VII, Factor VIII, von Willebrand factor, Factor IX, Factor X, Factor XI, C1 inhibitor, Protein C, and Protein S.
Fractionation workflows involving bound and non-bound material fractions using adsorption columns and elution buffers, with further processing of the non-bound material fraction to obtain non-bound proteins.
Workflows that include virus elimination steps as described in dependent claims and supported embodiments.
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