Granted Patent
B2
US 12,371,457 · App. 16/415,187 · Granted Jul 29, 2025
Protein expression strains
Inventors:
Christopher John Arthur Finnis (Nottingham, GB); Jennifer Mary Mclaughlan (Nottingham, GB); Per Kristoffer Nordeide (Copenhagen, DK); Preethi Ramaiya (Sunnyvale, CA)
Assignee:
Sartorius Albumedix Limited
C07K14/395C07K1/14C12N1/14C12N1/185C12N9/104C12N9/93C12N15/04C12N15/63C12N15/80C12N15/81C12P21/02C12Y603/02019C12R2001/865
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Abstract
The invention provides an improved host strain for production of desired protein.
Claims (41)
1. A fungal host cell comprising a modified NOT4 gene, wherein the Not4 protein encoded by the modified NOT4 gene comprises a mutation at a position corresponding to a position selected from the group consisting of: 429, 430, 434, and 437 of SEQ ID NO: 2, which results in:
a. a reduced level of expression of Not4 protein,
b. a reduced activity level of Not4 protein, or
c. a reduced level of expression of NOT4 gene
wherein the reduced level is relative to that of a wild-type Not4 protein or NOT4 gene of a reference fungal host cell,
wherein each of said fungal host cell and said reference fungal host cell is Saccharomyces cerevisiae,
wherein each of said fungal host cell and said reference fungal host cell comprises a nucleotide sequence encoding a heterologous protein which is an albumin or a fusion thereof,
wherein the heterologous protein has a higher yield in the fungal host cell than in the reference fungal host cell.
2. The fungal host cell of claim 1 , wherein the reference fungal host cell has a Not4 protein that comprises or consists of SEQ ID NO: 2.
3. The fungal host cell according to claim 1 in which the Not4 protein encoded by the modified NOT4 gene comprises a substitution from F to I at a position corresponding to position 429 of SEQ ID NO: 2.
4. The fungal host cell according to claim 3 , comprising a polynucleotide encoding SEQ ID NO: 4.
5. A method for producing a heterologous protein which is an albumin or fusion thereof from a fungal host cell comprising:
(i) providing a fungal host cell according to claim 1 , and
(ii) culturing the fungal host cell or culture to produce the desired protein.
6. A method for increasing the yield of a heterologous protein which is an albumin or fusion thereof comprising:
(i) providing a fungal host cell according to claim 1 , and
(ii) culturing the fungal host cell to produce the desired protein.
7. The method according to claim 5 in which the yield of the heterologous protein is at least 2% higher than the yield from a reference fungal host cell having the Not4 protein of SEQ ID NO: 2.
8. The method according to claim 5 in which the fungal host cell is cultured at a scale of at least 5 L.
9. The method according to claim 6 in which the yield of the heterologous protein is at least 2% higher than the yield from a reference fungal host cell having a Not4 protein of SEQ ID NO: 2.
10. The method according to claim 6 in which the fungal host cell is cultured at a scale of at least 5 L.
11. The method according to claim 5 , wherein the method further comprises:
(iii) recovering the heterologous protein,
(iv) purifying the heterologous protein,
(v) formulating the heterologous protein with a therapeutically acceptable carrier or diluent thereby to produce a therapeutic product suitable for administration to a human or an animal, and/or
(vi) providing the heterologous protein in unit dosage form.
12. The method according to claim 6 , wherein the method further comprises:
(iii) recovering the heterologous protein,
(iv) purifying the heterologous protein,
(v) formulating the heterologous protein with a therapeutically acceptable carrier or diluent thereby to produce a therapeutic product suitable for administration to a human or an animal, and/or
(vi) providing the heterologous protein in unit dosage form.
13. A method for producing a heterologous protein which is an albumin or fusion thereof from a fungal host cell comprising:
(i) providing a fungal host cell according to claim 3 , and
(ii) culturing the fungal host cell or culture to produce the heterologous protein.
14. The method according to claim 13 in which the yield of the heterologous protein is at least 2% higher than the yield from a reference fungal host cell having a Not4 protein of SEQ ID NO: 2.
15. The method according to claim 13 in which the fungal host cell is cultured at a scale of at least 5 L.
16. A method for producing a heterologous protein which is an albumin or fusion thereof from a fungal host cell comprising:
(i) providing a fungal host cell according to claim 4 , and
(ii) culturing the fungal host cell or culture to produce the heterologous protein.
17. The method according to claim 16 in which the yield of the heterologous protein is at least 2% higher than the yield from a reference fungal host cell having a Not4 protein of SEQ ID NO: 2.
18. The method according to claim 16 in which the fungal host cell is cultured at a scale of at least 5 L.
Assignments (5)
CHANGE OF NAME
Recorded Mar 11, 2025
From: ALBUMEDIX LTD
To: SARTORIUS ALBUMEDIX LIMITED
Reel/Frame 070468/0573 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Aug 12, 2022
From: ALBUMEDIX A/S
To: ALBUMEDIX LTD.
Reel/Frame 060791/0264 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Aug 8, 2022
From: FINNIS, CHRISTOPHER JOHN ARTHUR; MCLAUGHLAN, JENNIFER MARY; NORDEIDE, PER KRISTOFFER
To: ALBUMEDIX A/S
Reel/Frame 060746/0001 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Aug 8, 2022
From: NOVOZYMES A/S
To: ALBUMEDIX A/S
Reel/Frame 060746/0226 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Aug 8, 2022
From: RAMAIYA, PREETHI
To: NOVOZYMES A/S
Reel/Frame 061116/0673 →
Priority Claims (1)
EP 16152977
· Jan 27, 2016
· regional
Continuity (5)
Continuation
PCTUS2016068239
· Dec 22, 2016
Provisional Application
62278728
· Jan 14, 2016
Provisional Application
62270762
· Dec 22, 2015
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