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US4426323A Expired / Lapsed US ⏱️ 0.0 years remaining

Selected recovery of proteins from fermentation broths

Assignee: Veolia WTS Systems USA Inc
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Filing: 1981-06-08
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Granted: 1984-01-17
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Expires: 2001-06-08
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Abstract

Abstract A process is described for the selective recovery and/or concentration of a protein or a class of proteins from fermentation broth employing a combination of desalting and pH adjustment thereby causing the initial precipitation of unwanted proteins, again subjecting the resulting supernatant to another set of pH adjustment and desalting to cause the protein of interest to precipitate thereby obtaining a highly enriched fraction of the selected protein. Such a process will find use in the specific recovery of genetically engineered proteins from fermentation broths, e.g. interferon, insulin, growth hormone, etc.

Inventors

  • 👤 Surendar Jain

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-31 15:15
CPC Classifications
B; B01; B01D; B01D61/00; B01D61/42; B01D61/422; A; A23; A23C; A23C9/00; A23C9/14; A23C9/144; A23J; A23J1/00; A61; A61M; A61M1/00; A61M1/34; A61M1/3472; A61M1/3482; A61M1/3486; B01D2311/00; B01D2311/04; Y; Y10; Y10S; Y10S435/00; Y10S435/811

Claims Analysis

Total Claims: 15
Claim 1
Independent Claim 207 words
A process for separating and recovering a selected protein or selected group of proteins from a fermentation broth containing a mixture of selected and non-selected proteins comprising the steps of: (a) removing when present substantially all initial turbidity from said broth; (b) subsequently passing said broth into and out of a membrane desalting apparatus in contact with one surface of one or more semi-permeable membranes, said membrane separating said broth stream from an adjacent salt receiving stream thereby causing a decrease in the salt concentration of said broth; (c) said broth being adjusted in pH to within substantially the collective isoelectric point range of the non-selected protein sufficient to cause the formation of a first turbid phase containing essentially the said non-selected proteins and a first liquid phase enriched in the said selected protein; (d) subsequently removing substantially all of said first formed turbid phase; (e) passing the remaining first liquid phase into and out of said desalting apparatus to further decrease the salt concentration; (f) adjusting the pH to within the isoelectric point of the selected protein of interest thereby forming a second turbid phase enriched in the said selected protein of interest; (g) and separating the said second turbid phase for subsequent resuspension or purification.
Claim 2
Dependent Claim 207 words
A process for separating and recovering a selected protein or selected group of proteins from a fermentation broth containing a mixture of selected and non-selected proteins comprising the steps of: (a) removing when present substantially all initial turbidity from said broth; (b) subsequently passing said broth into and out of a membrane desalting apparatus in contact with one surface of one or more semi-permeable membranes, said membrane separating said broth stream from an adjacent salt receiving stream thereby causing a decrease in the salt concentration of said broth; (c) said broth being adjusted in pH to within substantially the collective isoelectric point range of the non-selected protein sufficient to cause the formation of a first turbid phase containing essentially the said non-selected proteins and a first liquid phase enriched in the said selected protein; (d) subsequently removing substantially all of said first formed turbid phase; (e) passing the remaining first liquid phase into and out of said desalting apparatus to further decrease the salt concentration; (f) adjusting the pH to within the isoelectric point of the selected protein of interest thereby forming a second turbid phase enriched in the said selected protein of interest; (g) and separating the said second turbid phase for subsequent resuspension or purification.
Claim 3
Dependent Claim 27 words
The process of claim 1 wherein the pH of the remaining first liquid phase is adjusted to between about 5-8 prior to further desalting in step (e).
Claim 4
Dependent Claim 198 words
A process for separating and recovering interferon from a fermentation broth containing a mixture of interferon and other proteins comprising the steps of: (a) removing when present substantially all initial turbidity from said broth; (b) subsequently passing said broth into and out of a membrane desalting apparatus in contact with one surface of one or more semi-permeable membranes, said membrane separating said broth stream from an adjacent salt receiving stream thereby causing the removal of between about 60-80% of the original salt concentration of said broth; (c) said broth being adjusted in pH to between about 4-5 to cause the formation of a first turbid phase containing essentially the said other proteins and a first liquid phase enriched in said interferon; (d) subsequently removing substantially all of said first formed turbid phase; (e) passing said remaining first liquid phase into and out of said desalting apparatus causing the further removal of at least about 90% of the original salt concentration of said broth; (f) adjusting the pH to within the isoelectric point of interferon thereby forming a second turbid phase enriched in said interferon; (g) and separating the said interferon enriched turbid phase for subsequent resuspension or purification.
Claim 5
Dependent Claim 198 words
A process for separating and recovering interferon from a fermentation broth containing a mixture of interferon and other proteins comprising the steps of: (a) removing when present substantially all initial turbidity from said broth; (b) subsequently passing said broth into and out of a membrane desalting apparatus in contact with one surface of one or more semi-permeable membranes, said membrane separating said broth stream from an adjacent salt receiving stream thereby causing the removal of between about 60-80% of the original salt concentration of said broth; (c) said broth being adjusted in pH to between about 4-5 to cause the formation of a first turbid phase containing essentially the said other proteins and a first liquid phase enriched in said interferon; (d) subsequently removing substantially all of said first formed turbid phase; (e) passing said remaining first liquid phase into and out of said desalting apparatus causing the further removal of at least about 90% of the original salt concentration of said broth; (f) adjusting the pH to within the isoelectric point of interferon thereby forming a second turbid phase enriched in said interferon; (g) and separating the said interferon enriched turbid phase for subsequent resuspension or purification.
Claim 6
Dependent Claim 26 words
The process of claim 3 wherein the pH of the remaining first liquid phase is adjusted to substantially neutral prior to further desalting in step (e).
Claim 7
Dependent Claim 13 words
A process of claim 1 wherein the membrane desalting apparatus is a dialyzer.
Claim 8
Dependent Claim 13 words
A process of claim 1 wherein the membrane desalting apparatus is an electrodialyzer.
Claim 9
Dependent Claim 21 words
A process of claim 1 wherein the pH adjustment in step (c) is made prior to said desalting of step (b).
Claim 10
Dependent Claim 22 words
A process of claim 1 wherein the pH adjustment in step (c) is made simultaneously with the said desalting of step (b).
Claim 11
Dependent Claim 21 words
A process of claim 1 wherein the pH adjustment in step (c) is made after the said desalting of step (b).
Claim 12
Dependent Claim 30 words
A process of claim 1 wherein the pH adjustment in step (c) is accomplished by controlling the pH of the said adjacent salt receiving stream of the membrane desalting apparatus.
Claim 13
Dependent Claim 22 words
A process of claim 1 wherein the pH adjustment in step (f) is made simultaneously with the said desalting of step (e).
Claim 14
Dependent Claim 21 words
A process of claim 1 wherein the pH adjustment in step (f) is made after the said desalting of step (e).
Claim 15
Dependent Claim 30 words
A process of claim 1 wherein the pH adjustment in step (f) is accomplished by controlling the pH of the said adjacent salt receiving stream of the membrane desalting apparatus.

References Cited (Backward Citations)

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USPTO Annuity & Maintenance Windows

Annuity Stage Window Opens Payment Deadline Surcharge Period Until Estimated Fee Status
3.5 Years (Year 4) 1987-01-16 1987-07-18 1988-01-17 $2000.00 Window Closed / Lapsed
7.5 Years (Year 8) 1991-01-16 1991-07-18 1992-01-17 $3760.00 Window Closed / Lapsed
11.5 Years (Year 12) 1995-01-16 1995-07-18 1996-01-17 $7700.00 Window Closed / Lapsed

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