Dac hyp compositions and methods

Patent metadata

Field	Value
Publication number	US-2020157230-A1
Application number	US-202016779336-A
Country	US
Kind code	A1
Filing date	Jan 31, 2020
Priority date	May 27, 2011
Publication date	May 21, 2020
Grant date	—

Abstract

The present disclosure relates to compositions of daclizumab suitable for subcutaneous administration and methods of manufacturing thereof.

First claim

What is claimed is:

1 . A modified NS0 cell that has been adapted to grow in serum- and cholesterol-free media and that is engineered to express a recombinant protein, said cell being capable of achieving a volumetric productivity exceeding 100 mg/L/day recombinant protein in a culture of 100 L in a 10-day fed-batch process when grown in serum- and cholesterol-free media.

2 . The modified NS0 cell of claim 1 which is capable of achieving a volumetric productivity exceeding:
(a) 100 mg/L/day recombinant protein in a culture of 1,000 L in a 10-day fed-batch process when grown in media free of cholesterol and animal-derived components;
(b) 100 mg/L/day recombinant protein in a culture of 16,000 L in a 10-day fed-batch process when grown in media free of cholesterol and animal-derived components;
(c) 200 mg/L/day recombinant protein in a culture of at least 100 L in a 13-day fed-batch process;
(d) 200 mg/L/day recombinant protein in a culture of 1,000 L in a 13-day fed-batch process when grown in cholesterol-free media; or
(e) 100 mg/L/day recombinant protein in a culture of 16,000 L in a 10-day fed-batch process when grown in serum- and cholesterol-free media.

3 . The modified NS0 cell of claim 1 , wherein said fed-batch process comprises adding a feed medium is added according to the following schedule, where the volume added represents the percentage of the initial cell culture volume:

Day
Volume added

0
0

1
0

2
4

3
7.8

4
7.8

5
7.8

6
11

7
13

8
15

9
15

10
0

4 . The modified NS0 cell of claim 1 which is stably transfected with a nucleic acid useful for expressing an anti-CD25 monoclonal antibody, optionally in which the anti-CD25 monoclonal antibody comprises a V L chain corresponding in sequence to positions 21-233 of SEQ ID NO:2 and a V H chain corresponding in sequence to positions 20 to 465 of SEQ ID NO:4.

5 . A method of producing a recombinant protein, comprising culturing the modified NS0 cell of claim 1 :
(a) under conditions that result in the production of at least 100 mg/L/day recombinant protein in a 100 L, 1,000 L or 16,000 L culture in a 10-day fed-batch process, or at least 200 mg/L/day recombinant protein in a 100 L, 1,000 L or 16,000 L culture in a 13-day fed-batch process; (b) in the absence of serum and cholesterol, and optionally in the absence of tropolone and hydrocortisone; (c) in a basal and/or feed medium containing 10-35 g/L glucose; or (d) in a basal medium containing 15 g/L glucose and/or a feed medium containing 28 g/L glucose, optionally wherein the basal medium is composed of the components of PFBM2±10% or PFFM3±10% and wherein the cell is cultured in basal medium for 1-3 days, and then in feed medium for 10-13 days.

6 . A vector useful for recombinantly expressing a protein of interest, comprising a weak promoter driving expression of a selectable marker operable in mammalian cells and a strong promoter driving expression of a protein of interest, optionally wherein the vector is pAbX.gpt or pHAT.IgG1.rg.dE and/or the protein of interest is (a) a therapeutic antibody; (b) an anti-CD25 antibody; (c) an anti-CD25 antibody which comprises the CDRs of daclizumab; or (d) daclizumab.

7 . A method for obtaining a mammalian host cell that has a high volumetric productivity of a protein of interest, comprising transfecting the cell with the vector of claim 6 , and selecting a cell that is capable of producing at least 100 mg/L/day protein of interest in a 100 L, 1,000 L or 16,000 L culture in a 10-day fed-batch process or at least 200 mg/L/day recombinant protein in a 100 L, 1,000 L or 16,000 L culture in a 13-day fed-batch process.

8 . A composition comprising daclizumab, wherein the daclizumab is characterized by the presence of a pE/Q heavy chain N-linked isoform and/or a Q/VHS heavy chain N-terminal isoform, optionally wherein the pE/Q heavy chain N-terminal isoform constitutes approximately 3-17%, approximately 3-15%, approximately 6-15%, approximately 5-15%, approximately 5-12%, or approximately 7-12% of the daclizumab, and, optionally wherein the Q/VHS heavy chain N-terminal isoform constitutes approximately 1-15% or approximately 3-12% of the daclizumab.

9 . The composition of claim 8 in which the heavy chain of daclizumab exists in the following N-terminal isoforms:

Isoform
Prevalence

(a)

pE/pE
25%-50% 

pE/Q
3%-15%

pE/VHS
25%-48% 

Q/VHS
1%-15%

VHS/VHS
0.5%-25%  

or (b)

pE/pE
 31-46%

pE/Q
5%-12%

pE/VHS
31%-42% 

Q/VHS
3%-12%

VHS/VHS
1%-17%

Assignees

• Abbvie Biotherapeutics Inc

Inventors

• Hartman Taymar E
• Sauer Paul W
• Burky John E
• Wesson Mark C
• Huang Ping Y
• Robinson Thomas J
• Partridge Braeden D
• Tso J Yun

Classifications

• A61K2039/505

  comprising antibodies · CPC title

• C07K2317/732

  Antibody-dependent cellular cytotoxicity [ADCC] · CPC title

• C07K2317/24

  containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered · CPC title

• C07K2317/76

  Antagonist effect on antigen, e.g. neutralization or inhibition of binding · CPC title

• A61K38/215

  IFN-beta · CPC title