EP0131789A2
Expired / Lapsed
EP
⏱️ 0.0 years remaining
Composition comprising purified human tumour necrosis factor in conjunction with human interferon and its use
Assignee: Memorial Sloan Kettering Cancer Center
•
Filing: 1984-06-23
•
Granted: —
•
Expires: 2004-06-23
Abstract
Abstract A method for the production and purification of human tumor necrosis factor(hTNF) is described.This purified material, shows growth inhibitory, hemorrhagic necrosis, cytotoxicity, and cytostasis effects on a variety of human tumors and cancers. The hTNF is free of exogeneous endotoxin, bacteria, and serum factors, hTNF has a molecular weight of about 70,000 hTNF and human interferon used together have a far greater growth inhibitory or cytotoxic effect on human tumors than the sum of their separate effects. The effect is synergistic and is seen with alpha-, beta- or gamma-IFN and with natural and recombinant IFN.
Inventors
- 👤 Lloyd J. Old
- 👤 Elizabeth Carswell Richards
- 👤 Berish Y. Rubin
- 👤 Barbara D. Williamson
- 👤 Jay S. Prendergast
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-10-24 20:34
CPC Classifications
C; C12; C12N; C12N15/00; C07; C07K; C07K14/00; C07K14/435; C07K14/52; C07K14/525; A; A61; A61K; A61K38/00; A61K38/16; A61K38/17; A61K38/19; A61K38/21
Claims Analysis
Total Claims: 45
Claim 1
Independent Claim
2 words
Purified hTNF.
Claim 2
Dependent Claim
2 words
Purified hTNF.
Claim 3
Dependent Claim
15 words
hTNF of Claim 1 having a molecular weight in the range of about 70,000 daltons.
Claim 4
Dependent Claim
14 words
hTNF of Claim 1 stable in the approximate pH range of 6 - 8.
Claim 5
Dependent Claim
28 words
hTNF of Claim 1 having a specific activity in the range of 2 - 5 x 10 units/mg Protein as determined by the mouse L-cell in vitro assay.
Claim 6
Dependent Claim
12 words
hTNF of Claim 1 stable at -78 C and -20 C. °
Claim 7
Dependent Claim
11 words
hTNF of Claim 1 stable at 56 C for 60 min.
Claim 8
Dependent Claim
50 words
Method for producing purified hTNF which comprises: (i) incubating human hematopoietic cells initially in tissue culture; (ii) separating the cell culture supernatant from said cells, (iii) collecting said cells, (iv) resuspending said cells and (v) re-incubating them; (vi) collecting the cell supernatant; and (vii) freezing the supernatant without said cells.
Claim 9
Dependent Claim
50 words
Method for producing purified hTNF which comprises: (i) incubating human hematopoietic cells initially in tissue culture; (ii) separating the cell culture supernatant from said cells, (iii) collecting said cells, (iv) resuspending said cells and (v) re-incubating them; (vi) collecting the cell supernatant; and (vii) freezing the supernatant without said cells.
Claim 10
Dependent Claim
27 words
Method of Claim 7 wherein the initial incubation of step (i) is in RPMI 1640 medium containing FCS in a range of about 0 - 8 %.
Claim 11
Dependent Claim
16 words
Method of Claim 7 wherein the cells are reincubated in step (iv) in RPMI 1640 medium.
Claim 12
Dependent Claim
12 words
Method of Claim 7 wherein the hematopoietic cells are human B cells.
Claim 13
Dependent Claim
40 words
Method of Claim 10 wherein the human B cells are selected from the group of BE, BI, CL, CW, DE, DM, DS, EJ, EL, EQ, ER, FC, FD, FG, ARH-77, DAUDI, LUKII, RPMI 1788, RPMI 8226, RPMI 8866 and ARA-10.
Claim 14
Dependent Claim
17 words
Method of Claim 7 wherein the cells are grown in step (i) in the presence of PMA.
Claim 15
Dependent Claim
59 words
Method of Claim 12, wherein after the initial incubation of step (i) said cells are resuspended in serum free R-ITS PREMIX and 2nM ethanolamine, the cell-free supernatants are collected and concentrated, said concentrated cell supernatant is contacted with a separation column equilibrated with buffer at pH 7.3 and the hTNF-active fractions obtained from said column are pooled and concentrated.
Claim 16
Dependent Claim
5 words
Composition comprising hTNF and hIFN.
Claim 17
Dependent Claim
5 words
Composition comprising hTNF and hIFN.
Claim 18
Dependent Claim
15 words
Composition of Claim 14 wherein the hTNF is produced by the method of Claim 7.
Claim 19
Dependent Claim
15 words
Composition of Claim 14 wherein the hTNF is produced by the method of Claim 12.
Claim 20
Dependent Claim
15 words
Composition of Claim 14 wherein the hTNF is produced by the method of Claim 13.
Claim 21
Dependent Claim
20 words
Composition of Claim 14 wherein the hIFN is selected from the group of alpha-hIFN, beta-hIFN, gamma hIFN and mixtures thereof.
Claim 22
Dependent Claim
10 words
Composition of Claim 14 wherein the hIFN is natural hIFN.
Claim 23
Dependent Claim
10 words
Composition of Claim 14 wherein the hIFN is recombinant hIFN.
Claim 24
Dependent Claim
27 words
Method for distinguishing normal from malignant cells in a mixture which comprises contacting said cells with hTNF and observing a growth inhibitory effect on the malignant cells.
Claim 25
Dependent Claim
27 words
Method for distinguishing normal from malignant cells in a mixture which comprises contacting said cells with hTNF and observing a growth inhibitory effect on the malignant cells.
Claim 26
Dependent Claim
17 words
Method of Claim 21 wherein hTNF is produced according to the process of Claims 7 - 13.
Claim 27
Dependent Claim
32 words
Method of disthinguishing normal from malignant cells in a mixture which comprises contacting said cells with a composition comprising hTNF and hIFN and observing a growth inhibitory effect on the malignant cells.
Claim 28
Dependent Claim
32 words
Method of disthinguishing normal from malignant cells in a mixture which comprises contacting said cells with a composition comprising hTNF and hIFN and observing a growth inhibitory effect on the malignant cells.
Claim 29
Dependent Claim
17 words
Method of Claim 23 wherein hTNF is produced according to the process of Claims 7 - 13.
Claim 30
Dependent Claim
19 words
Method of Claim 23 wherein the hIFN is selected from the group of alpha-hIFN, beta-hIFN, gamma-hIFN and mixtures thereof.
Claim 31
Dependent Claim
10 words
Method of Claim 25 wherein the hIFN is natural hIFN.
Claim 32
Dependent Claim
10 words
Method of Claim 25 wherein the hIFN is recombinant hIFN.
Claim 33
Dependent Claim
20 words
Method for producing mouse L(R) cells resistant to hTNF by repeated passage of mouse L(S) cells through medium containing hTNF.
Claim 34
Dependent Claim
20 words
Method for producing mouse L(R) cells resistant to hTNF by repeated passage of mouse L(S) cells through medium containing hTNF.
Claim 35
Dependent Claim
17 words
Method of Claim 28 wherein the hTNF is produced by the method of Claims 7 - 13.
Claim 36
Dependent Claim
26 words
Method for inhibiting growth of human cancer cells which comprises exposing the human cancer cells to amounts of hTNF sufficient to inhibit growth of said cells.
Claim 37
Dependent Claim
26 words
Method for inhibiting growth of human cancer cells which comprises exposing the human cancer cells to amounts of hTNF sufficient to inhibit growth of said cells.
Claim 38
Dependent Claim
17 words
Method of Claim 30 wherein the hTNF is produced by the method of Claims 7 - 13.
Claim 39
Dependent Claim
30 words
Method for inhibiting growth of human cancer cells which comprises exposing the human cancer cells to a composition comprising of hTNF and hIFN sufficient to inhibit growth of said cells.
Claim 40
Dependent Claim
30 words
Method for inhibiting growth of human cancer cells which comprises exposing the human cancer cells to a composition comprising of hTNF and hIFN sufficient to inhibit growth of said cells.
Claim 41
Dependent Claim
17 words
Method of Claim 32 wherein hTNF is produced according to the process of Claims 7 - 13.
Claim 42
Dependent Claim
19 words
Method of Claim 32 wherein the hIFN is selected from the group of alpha-hIFN, beta-hIFN, gamma-hIFN and mixtures thereof.
Claim 43
Dependent Claim
10 words
Method of Claim 34 wherein the hIFN is natural hIFN.
Claim 44
Dependent Claim
10 words
Method of Claim 34 wherein the hIFN is recombinant hIFN.
Claim 45
Dependent Claim
13 words
Purified hTNF of claim 1 as produced by the method of claims 7-13.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
Maintenance schedule is calculated for granted US utility patents.