1. A synchronized controlled oscillation modulator (SCOM), comprising:
at least one controlled oscillation modulator (COM) having a non-hysteresis comparator, a switching stage and a negative feedback loop adapted to achieve conditions for controlled oscillation of a predetermined frequency by means of at least two poles, and
a synchronizing device connected to said COM modulator.
2. A modulator according to claim 1, wherein the synchronizing device comprises an oscillation signal generator.
3. A modulator according to claim 1, wherein the modulator is synchronized by another SCOM modulator comprising any oscillating modulator signal with the frequency of a wanted idle switching frequency.
4. A modulator according to claim 1, comprising a second COM modulator, said synchronizing device being connected between forward paths of said COM modulators and arranged to synchronize the COM modulators with each other.
5. A modulator according to claim 3, wherein said synchronization device comprises a circuit consisting of R, RC or C components.
6. A modulator according to claim 5, wherein said synchronizing device comprises:
a first and a second series resistances (RA, RB), a parallel resistor (Rosc), and a parallel capacitor (Cosc), wherein said first series resistance is connected to a first end of the parallel capacitor and parallel resistor, and said second series resistance is connected to a second end of the parallel capacitor and parallel resistor, and wherein said first and second series resistances are connected to a comparator in the forward path of each modulator respectively.
7. A modulator according to claim 3, wherein said-synchronization-device comprises an active circuit, preferably including at least one high pass filter.
8. A modulator according to claim 1, wherein the modulator is extended by an additional SCOM modulator driven in a full-bridge configuration to achieve a three-level pulse output.
9. A modulator according to claim 8, wherein the modulator is implemented in a PSCPWM system, without common mode high frequency spectral contributions in the three level pulse output.
10. A modulator according to claim 1, wherein the modulator is implemented in a multi-loop MECC (N,M) control system for enhanced noise suppression, where N and M are integers representing the number of local and global control loops respectively.
11. A modulator according to claim 1, further comprising N COM modulators synchronized by an additional synchronization signal or by a common COM signal of said COM modulators.
12. A modulator according to claim 1, further comprising a limiter device to control the PWM modulation depth.
13. A modulator according to claim 1, wherein the modulator is implemented in a general power conversion system, in the general power conversion system is in a DC-AC audio power conversion system.
14. A modulator according to claim 1, wherein the modulator is used to drive an electrodynamic transducer load directly.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A process for the preparation of Neutrophil Inhibitory Factor comprising the step of growing a cell line expressing Neutrophil Inhibitory Factor in an animal component-free medium selected from the group consisting of an inoculum growth medium, a production growth medium and a nutrient feed to give a production culture.
2. A process according to claim 1 wherein the Neutrophil Inhibitory Factor is the protein of SEQ. ID. NO. 3.
3. A process for the according to claim 2 wherein the protein is glycosylated and has a relative molecular weight of about 38.3 to about 64.1 kDa.
4. A process according to claim 2 wherein the protein is about 5 to about 25% mono-sialylated; about 10 to about 30% di-sialylated, about 15 to about 35% tri-sialylated, about 15 to about 45% tetra-sialylated and about 1 to about 20% non-sialylated.
5. A process according to claim 1 wherein the animal component-free production growth medium comprises:
(i) a CHO-III-PFMglucose solution;
(ii) a sodium hypoxanthinethymidine solution; and
(iii) yeast extract.
6. A process according to claim 1 wherein the animal component-free production growth medium comprises:
(i) CHO-III-PFMglucose solution;
(ii) about 5 to about 20 ml per liter (i) of a 10 mM sodium hypoxanthine1.6 mM thymidine solution; and
(iii) about 0.5 to about 5.0 grams per liter (i) yeast extract.
7. A process according to claim 1 wherein the animal component-free production growth medium comprises:
(i) CHO-III-PFMglucose;
(ii) 10.0 ml per liter (i) of a 10 mM sodium hypoxanthine1.6 mM thymidine solution; and
(iii) 1.5 grams per liter (i) yeast extract.
8. A process according to claim 1 further comprising the steps of:
(a) providing an inoculum prepared by incubating a cell line expressing Neutrophil Inhibitory Factor in an animal component-free inoculum growth medium; and
(b) transferring said inoculum to a vessel containing an animal component-free production growth medium.
9. A process according to claim 8 wherein the inoculum growth medium comprises:
(i) a CHO-III-PFMglucose;
(ii) a sodium hypoxanthinethymidine solution;
(iii) an amino acid solution comprising acids selected from the group consisting of L-aspartic acid, L-glutamic acid, L-asparagine, L-proline, L-serine, and L-methionine;
(iv) optionally an L-methionine sulphoximine solution; and
(v) an L-cysteine solution.
10. A process according to claim 8 wherein the inoculum growth medium comprises:
(i) CHO-III-PFMglucose solution;
(ii) about 5 to about 20 ml per liter (i) of a 10 mM sodium hypoxanthine1.6 mM thymidine solution;
(iii) about 5 to about 30 ml per liter (i) of an amino acid solution comprising L-aspartic acid (3.0 gl), L-glutamic acid (2.5 gl), L-asparagine (10.0 gl), L-proline (1.25 gl), L-serine (3.0 gl), and L-methionine (1.5 gl);
(iv) about 0 to about 75 mol per liter (i) of L-methionine sulphoximine; and
(v) about 10 to about 40 mg per liter (i) of L-cysteine.
11. A process according to claim 8 wherein the inoculum growth medium comprises:
(i) CHO-III-PFMglucose;
(ii) 10.0 ml per liter (i) of a 10 mM sodium hypoxanthine1.6 mM thymidine solution;
(iii) 20.0 ml per liter (i) of an amino acid solution comprising L-aspartic acid (3.0 gl), L-glutamic acid 2.5 gl), L-asparagine (10.0 gl), L-proline (1.25 gl), L-serine (3.0 gl), and L-methionine (1.5 gl);
(iv) optionally 1.0 ml per liter (i) of a 25 mM L-methionine sulphoximine solution; and
(v) 25.0 mg per liter (i) of L-cysteine.
12. A process according to claim 8 further comprising the step:
(c) feeding the production culture with at least one nutrient feed.
13. A process according to claim 12 wherein step (c) includes a first nutrient feed and a second nutrient feed.
14. A process according to claim 13 wherein the first nutrient feed is a nutrient feed comprising an aqueous solution of about 100 to about 500 grams of glucose per liter.
15. A process according to claim 13 wherein the first nutrient feed is a nutrient feed comprising an aqueous solution of about 200 grams of glucose per liter.
16. A process according to claim 15 wherein the first nutrient feed is added at a rate of about 0.0 to about 6.0 grams of glucose per liter growth medium per day.
17. A process according to claim 13 wherein the second nutrient feed comprises
(i) a CHO-III-PFM (5) solution with 1 L-cystine, 3 L-tyrosine and without glucose, hypoxanthine, thymidine, L-glutamine, sodium bicarbonate, sodium chloride;
(ii) 25 to 100 ml per liter of solution (i) of a 10 mM sodium hypoxanthine1.6 mM thymidine solution; and
(iii) 5 to 20 grams per liter of solution (i) yeast extract.
18. A process according to claim 13 wherein the second nutrient feed comprises
(i) CHO-III-PFM (5) solution with 1 L-cystine, 3 L-tyrosine and without glucose, hypoxanthine, thymidine, L-glutamine, sodium bicarbonate, sodium chloride;
(ii) 50 ml per liter of solution (i) of a 10 mM sodium hypoxanthine1.6 mM thymidine solution; and
(iii) 7.5 grams per liter of solution (i) yeast extract.
19. A process according to claim 18 wherein the second nutrient feed is fed to the reactor continuously at a rate of approximately 25 mlliter-day.
20. A process according to claim 13 wherein said first nutrient feed comprises about 100 to about 500 grams per liter glucose and said second nutrient feed comprises (1) a CHO-III-PFM (5) solution, (2) about 25 to about 100 ml per liter (1) of a 10 mM sodium hypoxanthine1.6 mM thyandine solution; and (3) about 5 to about 5 grams per liter (1) yeast extract.
21. A process according to claim 12 wherein the nutrient feed is a nutrient feed comprising an aqueous solution of about 100 to about 500 grams of glucose per liter.
22. A process according to claim 12 wherein the nutrient feed is a nutrient feed comprising an aqueous solution of about 200 grams of glucose per liter.
23. A process according to claim 22 wherein the nutrient feed is added at a rate of about 0.0 to about 6.0 grams of glucose per liter growth medium per day.
24. A neutrophil inhibitory factor made by the process of claim 1.
25. An animal component-free production growth medium comprising:
(i) a CHO-III-PFMglucose solution;
(ii) a sodium hypoxanthinethymidine solution; and
(iii) yeast extract.
26. A medium according to claim 25 comprising:
(i) CHO-III-PFMglucose solution;
(ii) about 5 to about 20 ml per liter (i) of a 10 mM sodium hypoxanthine1.6 mM thymidine solution; and
(iii) about 0.5 to about 5.0 grams per liter (i) yeast extract.
27. A medium according to claim 25 comprising:
(i) CHO-III-PFMglucose solution;
(ii) 10.0 ml per liter (i) of a 10 mM sodium hypoxanthine1.6 mM thymidine solution; and
(iii) 1.5 grams per liter (i) yeast extract.
28. A method for the preparation of recombinant proteins comprising the cultivation of mammalian cells expressing an exogenous recombinant protein in the animal component-free growth medium of claim 25.
29. A method according to claim 28 wherein the mammalian cells are Chinese Hamster Ovary cells transfected with a glutamine synthetase plasmid vector containing the DNA coding region for the recombinant protein.
30. A method according to claim 29 wherein the vector is a glutamine synthetasemethionine sulfoximine co-amplification vector selected from pEE14 and pEE14.1.
31. An inoculum growth medium comprising:
(i) a CHO-III-PFMglucose solution;
(ii) a sodium hypoxanthinethymidine solution;
(iii) an amino acid solution comprising acids selected from the group consisting of L-aspartic acid, L-glutamic acid, L-asparagine, L-proline, L-serine, and L-methionine;
(iv) optionally an L-methionine sulphoximine solution; and
(v) an L-cysteine solution.
32. An inoculum growth medium according to claim 31 comprising:
(i) CHO-III-PFMglucose solution;
(ii) about 5 to about 20 ml per liter (i) of a 10 mM sodium hypoxanthine1.6 mM thymidine solution;
(iii) about 5 to about 30 ml per liter (i) of an amino acid solution comprising L-aspartic acid (about 3.0 gl), L-glutamic acid (about 2.5 gl), L-asparagine (about 10.0 gl), L-proline (about 1.25 gl), L-serine (about 3.0 gl), and L-methionine (about 1.5 gl);
(iv) about 0 to about 75 mol per liter (i) of an L-methionine sulphoximine; and
(v) about 10 to about 40 mg per liter (i) of L-cysteine.
33. An inoculum growth medium according to claim 32 comprising:
(i) CHO-III-PFMglucose solution;
(ii) 10.0 ml per liter (i) of a 10 mM sodium hypoxanthine1.6 mM thymidine solution;
(iii) 20.0 ml per liter (i) of an amino acid solution comprising L-aspartic acid (3.0 gl), L-glutamic acid 2.5 gl), L-asparagine (10.0 gl), L-proline (1.25 gl), L-serine (3.0 gl), and L-methionine (1.5 gl);
(iv) optionally 1.0 ml per liter (i) of a 25 mM L-methionine sulphoximine solution; and
(v) 25.0 mg per liter (i) of L-cysteine.
34. A nutrient feed comprising
(i) a CHO-III-PFM (5) solution with 1 L-cystine, 3x L-tyrosine and without glucose, hypoxanthine, thymidine, L-glutamine, sodium bicarbonate, sodium chloride;
(ii) 25 to 111 ml per liter of solution (i) of a 10 mM sodium hypoxanthine1.6 mM thymidine solution; and
(iii) 5 to 20 grams per liter of solution (i) yeast extract.
35. The cell line PFG-01 (ATCC PTA-2503).
36. A method for the preparation of Neutrophil Inhibitory Factor comprising culturing the cell line of claim 35 under conditions promoting expression of Neutrophil Inhibitory Factor and recovering the Neutrophil Inhibitory Factor.
37. A method according to claim 36 wherein the Neutrophil Inhibitory factor comprises the amino acid sequence of SEQ. ID. NO. 3.
38. A Neutrophil Inhibitory Factor prepared by culturing the cell line PFGOl (ATCC PTA-2503).
39. A Neutrophil Inhibitory Factor prepared by the method of claim 36 or 37.
40. A process according to any of claims 1 to 23 wherein said cell line is PFG01 (ATCC PTA-2503).
41. A Neutrophil Inhibitory Factor made by the process of any of claims 2 to 23.
42. An isolated Neutrophil Inhibitory Factor having neutrophil inhibitory activity and comprising the amino acid sequence of SEQ. ID. NO. 3 that is produced by cell line PFG01 (ATCC PTA-2503).