Process For Isolation of Plasma or Serum Proteins

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.

1 - 18 . (canceled) 
     
     
         19 : A process for the large-scale isolation of proteins from a protein solution wherein the protein solution is obtained from a source selected from the group consisting of human blood and a solution derived from human blood, the process comprising the steps of:
 a) adjusting the pH of the protein solution to a pH in the range of 3 to 10;   b) 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;   c) applying said protein solution to an adsorption column comprising an adsorbent, said adsorbent comprises a particle with at least one high density non-porous core, surrounded by a porous material, the adsorbent comprises a particle density of at least 1.5 g/ml and a mean volume particle diameter of at most 150 μm, whereby one or more proteins are bound to the adsorbent;   d) obtaining a non-bound material fraction from the column, which non-bound material fraction contains at least one non-bound protein;   e) subjecting the adsorbent to at least one elution buffer to elute at least one of the proteins bound to the adsorbent, thereby obtaining a bound material fraction; and   f) subjecting the non-bound material fraction to further processing to obtain at least one non-bound protein therefrom.   
     
     
         20 : The process according to  claim 19 , wherein the protein solution has been supplemented with an alcohol. 
     
     
         21 : The process according to  claim 19 , wherein the proteins are human blood proteins. 
     
     
         22 : The process according to  claim 20 , wherein after supplementation with alcohol the protein solution comprises a supernatant and a precipitated fraction. 
     
     
         23 : The process according to  claim 22 , wherein the supernatant and the precipitated fraction are separated by filtration, microfiltration, centrifugation, decantation, and/or sedimentation. 
     
     
         24 : The process according to  claim 23 , wherein the proteins are found in the supernatant. 
     
     
         25 : The process according to  claim 23 , wherein the proteins are found in the precipitated fraction. 
     
     
         26 : The process according to  claim 22 , wherein the supernatant or the precipitated fraction has a concentration of alcohol of at least 0.1% by volume. 
     
     
         27 : The process according to  claim 22 , wherein the alcohol is selected from the group consisting of methanol, ethanol, n-propanol, i-propanol, n-butanol, i-butanol, s-butanol, t-butanol, methylene glycol, ethylene glycol, propylene glycol, diethylene glycol, methylene-ethylene glycol and dimethylene glycol. 
     
     
         28 : The process according to  claim 22 , wherein the precipitated fraction is a resolubilised precipitate obtained by the addition of alcohol to human blood, or to a solution derived from human blood. 
     
     
         29 : The process according to  claim 22 , wherein the protein solution is obtained by re-combination of one or more supernatants and/or one or more resolubilised precipitates obtained by the stepwise addition of alcohol to human blood or a solution derived from human blood. 
     
     
         30 : The process according to  claim 20 , wherein the protein solution is obtained by a Cohn fractionation method or a variation thereof. 
     
     
         31 : The process according to  claim 19 , further comprising at least one virus elimination treatment. 
     
     
         32 : The process according to  claim 31 , wherein the at least one virus elimination treatment is performed prior to contacting the protein solution with the adsorbent. 
     
     
         33 : The process according to  claim 31 , wherein the virus elimination treatment involves addition of detergent and/or an organic solvent to the protein solution. 
     
     
         34 : The process according to  claim 19 , wherein the adsorbent comprises a functionalized matrix polymer carrying a plurality of covalently attached functional groups comprising an aromatic or heteroaromatic ring-system and/or one or more acidic groups. 
     
     
         35 : The process according to  claim 21 , wherein the human blood proteins are selected from the group consisting of albumin, IgG, IgA, IgM, IgD, IgE, alpha-1-proteinase inhibitor, blood pro-coagulation protein, blood anti-coagulation protein, thrombolytic agent, anti-angiogenic protein, α-2-antiplasmin, C-1 esterase inhibitor, apolipoprotein, HDL, LDL, Fibronectin, beta-2-glycoprotein I, fibrinogen, plasminogen, plasmin, plasminogen activator, plasminogen inhibitor, plasma protease inhibitor, anti-thrombin III, streptokinase, inter-alpha-trypsin inhibitor, α-2-macroglobulin, amyloid protein, ferritin, pre-albumin, GC-globulin, haemopexin, C3-complement, transferrin, urokinase, α-1-acid-glycoprotein, Factor II, Factor V, Factor VII, Factor VIII, von Willebrand factor, Factor VIII—von Willebrand factor complex, Factor IX, Factor X, Factor XI, C1-inhibitor, protein C and protein S. 
     
     
         36 : The process according to  claim 19 , wherein, after applying the protein solution to the adsorption column, one or more proteins are washed out as a non-bound protein with one or more washing buffers. 
     
     
         37 : The process according to  claim 19 , wherein one non-bound protein is an alpha-1-proteinase inhibitor. 
     
     
         38 - 66 . (canceled) 
     
     
         67 : A process for the large-scale isolation of proteins from a protein solution, wherein the protein solution is obtained from a source selected from the group consisting of human blood and a solution derived from human blood, the process comprising the steps of:
 a) providing a protein solution comprising one or more proteins and obtained from a source selected from the group consisting of human blood and a solution derived from human blood, the protein solution having a preset pH and a preset ionic strength or conductivity;   b) contacting the protein solution with an adsorbent, wherein the adsorbent comprises 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;   c) subjecting the adsorbent to an elution buffer to elute at least one of the proteins bound to the adsorbent; and   d) subjecting the adsorbent to one or more additional elution buffers to elute one or more remaining proteins bound to the adsorbent.   
     
     
         68 - 90 . (canceled)