1. A process for coating a substrate, comprising
producing a bimodal polyethylene composition by
subjecting ethylene, optionally with hydrogen andor comonomers to polymerization or copolymerization reactions in a multistage polymerization sequence of successive polymerization stages;
carrying out the polymerization reactions in the presence of a single site catalyst capable of forming a composition comprising
a low molecular weight component with an MFR2 of 20 g10 min or more and a density higher than the density of the composition, and
a high molecular weight component,
said composition having a melt flow rate MFR2 of 10\u201320g10 min and a density of 915\u2013960 kgm3; and
extruding said composition on the substrate as such or by adding 10 wt % or less, calculated from the total weight of the coating, of high pressure PE-LD by blending into the extrusion composition or by co-extrusion
wherein the active complex of said single site catalyst has the general formula
(X1)(X2)Hf(Cp-R1)(Cp-R2),
where X1 and X2 are thee same or different and selected from the group comprising halogen, methyl, benzyl, and hydrogen;
Hf is hafnium;
Cp is cyclopentadienyl group; and
R1 and R2 are same or different and stand for linear and branched hydrocarbyl groups containing 1\u201310 carbon atoms.
2. The process according to claim 1, wherein the bimodal polyethylene composition is produced by
subjecting ethylene, optionally with hydrogen andor comonomers to a first polymerization or copolymerization reaction in the presence of a single-site catalyst in a first reaction zone or reactor to produce a first polymerization product with an MFR2 of 20 g10 min or more;
recovering the first polymerization product from the first reaction zone;
feeding the first polymerization product to a second reaction zone or reactor;
feeding additional ethylene and, optionally, comonomers to the second reaction zone;
subjecting the additional ethylene and optionally additional monomer(s) andor hydrogen to a second polymerization reaction in the presence of the single-site catalyst and the first polymerization product to produce a second polymerization product having a MFR2 of 10\u201320 g10 min; and
recovering the combined polymerization product from the second reaction zone.
3. The process according to claim 1, wherein the substrate is paper or paperboard.
4. The process according to claim 1, wherein the active complex of the catalyst is bis-(n-butyl cyclopentadienyl) hafnium dihalide.
5. The process according to claim 1, wherein essentially no fresh catalyst is added to the reactors other than the first one.
6. The process according to claim 1, wherein the process is carried out in a polymerization reactor cascade comprising a loop reactor and a gas phase reactor, in that order.
7. The process according to claim 1, wherein the process is carried out in a polymerization reactor cascade comprising two or more gas phase reactors.
8. The process according to claim 1 or 2, wherein 0.1\u20135 wt-%, calculated from the total weight of the coating composition of high pressure PE-LD is blended into polyethylene composition prior to the extrusion.
9. The process according to claim 1 or 2, wherein 0.1\u20133 wt-%, calculated from the total weight of the coating composition of high pressure PE-LD is blended into the polyethylene composition prior to the extrusion.
10. A process for coating a substrate, comprising:
producing a bimodal polyethylene composition by
subjecting ethylene, optionally with hydrogen andor comonomers to polymerization or copolymerization reactions in a multistage polymerization sequence of successive polymerization stages;
carrying out the polymerization reactions in the presence of a single site catalyst capable of forming a composition comprising
a low molecular weight component with an MFR2 of 20 g10 min or more and a density higher than the density of the composition, and
a high molecular weight component,
said composition having a melt flow rate MFR2 of 10\u201320 g10 min and a density of 915\u2013960 kgm3;
extruding said composition on the substrate as such or by adding 10 wt % or less calculated from the total weight of the coating, of high pressure PE-LD by blending into the extrusion composition or by co-extrusion,
wherein the active complex of the catalyst is a siloxy-substituted bis-indenyl zirconium dihalide.
11. The process according to claim 10, wherein a metallocene catalyst is used as the single site catalyst.
12. A process for coating a substrate, comprising:
producing a bimodal polyethylene composition by
subjecting ethylene, optionally with hydrogen andor comonomers to polymerization or copolymerization reactions in a multistage polymerization sequence of successive polymerization stages;
carrying out the polymerization reactions in the presence of a single site catalyst capable of forming a composition comprising
a low molecular weight component with an MFR2 of 20 g10 min or more and a density higher than the density of the composition, and
a high molecular weight component,
said composition having a melt flow rate MFR2 of 10\u201320 g10 min and a density of 915\u2013960 kgm3; and
extruding said composition on the substrate as such or by adding 10 wt % or less, calculated from the total weight of the coating, of high pressure PE-LD by blending into the extrusion composition or by co-extrusion,
wherein 0.1\u20138 wt %, calculated from the total weight of the coating composition, of high pressure PE-LD is blended into the polyethylene composition prior to the extrusion,
wherein 30 to 70% of the ethylene homopolymer or copolymer is produced at conditions which provide a polymer having a MFR2 of 10 g10 min or more and 70% of the ethylene homopolymer or copolymer is produced at conditions which provide a polymer having a MFR2 of less than 10 g10 min or more and 70 to 30% of the ethylene homopolymer or copolymer is produced at conditions which provide a polymer having a MFR2 of less than 5 g10 min.
13. The process according to claim 10, wherein 40 to 60% of the ethylene homopolymer or copolymer is produced at conditions which provide a polymer having a MFR2 of 10 g10 min or more and 60 to 40% of the ethylene homopolymer or copolymer is produced at conditions which provide a polymer having a MFR2 of less than 5 g10 min.
14. The process according to claim 1 or 2, wherein the composition is coextruded on the substrate together with high pressure PE-LD so as to form a coating comprising 0.1\u20138 wt-%, calculated from the total weight of the coating, of high pressure PE-LD.
15. The process according to claim 14, wherein said coating composition comprises 0.1\u20135 wt-%, calculated from the total weight of the coating of high pressure PE-LD.
16. The process according to claim 14, wherein said coating composition comprises 0.1\u20133 wt-%, calculated from the total weight of the coating of high pressure PE-LD.
17. A process for coating a substrate, comprising
producing a bimodal polyethylene composition by
subjecting ethylene, optionally with hydrogen andor comonomers to polymerization or copolymerization reactions in a multistage polymerization sequence of successive polymerization stages;
carrying out the polymerization reactions in the presence of a single site catalyst capable of forming a composition comprising
a low molecular weight component with an MFR2 of 20 g10 min or more and a density higher than the density of the composition, and
a high molecular weight component, said composition having a melt flow rate MFR2 of 10\u201320 g10 min and a density of 915\u2013960 kgm3;
extruding said composition on the substrate as such or by adding 10 wt % or less, calculated from the total weight of the coating, of high pressure PE-LD by blending into the extrusion composition or by co-extrusion,
wherein the active complex of the catalyst is a siloxy-substituted bis-indenyl zirconium dihalide.
18. A process for coating a substrate, comprising
producing a bimodal polyethylene composition by
subjecting ethylene, optionally with hydrogen andor comonomers to polymerization or copolymerization reactions in a multistage polymerization sequence of successive polymerization stages;
carrying out the polymerization reactions in the presence of a single site catalyst capable of forming a composition comprising
a low molecular weight component with an MFR2 of 20 g10 min or more and a density higher than the density of the composition, and
a high molecular weight component, said composition having a melt flow rate MFR2 of 10\u201320 g10 min and a density of 915\u2013960 kgm3;
extruding said composition on the substrate as such or by adding 10 wt % or less, calculated from the total weight of the coating, of high pressure PE-LD by blending into the extrusion composition or by co-extrusion,
wherein 0.1\u20130.8 wt %, calculated from the total weight of the coating composition, of high pressure PE-LD is blended into the polyethylene composition prior to the extrusion,
wherein 30\u201370% of the ethylene homopolymer or copolymer is produced at conditions which provide a polymer having a MFR2 of 10 g10 min or more and 70\u201330% of the ethylene homopolymer or copolymer is produced at conditions which provide a polymer having a MFR2 of less than 5 g10 min;
wherein the active complex of said single site catalyst has the general formula
(X1)(X2)Hf(Cp-R1)(Cp-R2),
where X1 and X2 are thee same or different and selected from the group comprising halogen, methyl, benzyl, and hydrogen;
Hf is hafnium;
Cp is cyclopentadienyl group; and
R1 and R2 are same or different and stand for linear and branched hydrocarbyl groups containing 1\u201310 carbon atoms. R1 and R2 are same or different and stand for linear and branched hydrocarbyl groups containing 1\u201310 carbon atoms.
19. The process according to claim 18, wherein the substrate is paper or paperboard.
20. A process for coating a substrate, comprising
producing a bimodal polyethylene composition by
subjecting ethylene, optionally with hydrogen andor comonomers to polymerization or copolymerization reactions in a multistage polymerization sequence of successive polymerization stages;
carrying out the polymerization reactions in the presence of a single cite catalyst capable of forming a composition comprising
a low molecular weight component with a MFR2 of 20 g10 min or more and a density higher than 965 kgm3, but higher than the density of the composition, and
a high molecular weight component, said composition having a melt flow rate MFR2 of 10\u201320 g10 min and a density of 940\u2013960 kgm3; and
extruding said composition on the substrate as such or by adding 10 wt-% or less, calculated from the total weight of the coating, of high pressure PE-LD by blending into the extrusion composition or by co-extrusion
wherein the active complex of said single site catalyst has the general formula
(X1)(X2)Hf(Cp-R1)(Cp-R2),
where X1 and X2 are thee same or different and selected from the group comprising halogen, methyl, benzyl, and hydrogen;
Hf is hafnium;
Cp is cyclopentadienyl group; and
R1 and R2 are same or different and stand for linear and branched hydrocarbyl groups containing 1\u201310 carbon atoms.
21. The process according to claim 20, wherein the bimodal polyethylene composition is produced by
subjecting ethylene, optionally with hydrogen andor comonomers to a first polymerization or copolymerization reaction in the presence of a single-site catalyst in a first reaction zone or reactor to produce a first polymerization product with an MFR2 of 20 g10 min or more;
recovering the first polymerization product from the first reaction zone;
feeding the first polymerization product to a second reaction zone or reactor;
feeding additional ethylene and, optionally, comonomers to the second reaction zone;
subjecting the additional ethylene and optionally additional monomer(s) andor hydrogen to a second polymerization reaction in the presence of the single-site catalyst and the first polymerization product to produce a second polymerization product having a MFR2 of 5 g10 min or more; and
recovering the combined polymerization product from the second reaction zone.
22. The process according to claim 20 or 21, wherein said composition has a density of 915\u2013930 kgm3, and the low molecular weight component has a MFR2 of 20\u2013500 g10 min and a density of 935\u2013965 kgm3.
23. A process for coating a substrate, comprising:
producing a bimodal polyethylene composition by
subjecting ethylene, optionally with hydrogen andor comonomers to polymerization or copolymerization reactions in a multistage polymerization sequence of successive polymerization stages;
carrying out the polymerization reactions in the presence of a single cite catalyst capable of forming a composition comprising
a low molecular weight component with an MFR2 of 70\u20131000 g10 min or more and a density higher than 965 kgm3, and
a high molecular weight component, said composition having a melt flow rate MFR2 of 10\u201320 g10 min and a density of 940\u2013960 kgm3; and
extruding said composition on the substrate as such or by adding 10 wt-% or less, calculated from the total weight of the coating, of high pressure PE-LD by blending into the extrusion composition or by co-extrusion;
wherein the active complex of said single site catalyst has the general formula
(X1)(X2)Hf(Cp-R1)(Cp-R2),
where X1 and X2 are thee same or different and selected from the group comprising halogen, methyl, benzyl, and hydrogen;
Hf is hafnium;
Cp is cyclopentadienyl group; and
R1 and R2 are same or different and stand for linear and branched hydrocarbyl groups containing 1\u201310 carbon atoms.
24. A process for coating a substrate, comprising
producing a bimodal polyethylene composition by
subjecting ethylene, optionally with hydrogen andor comonomers to polymerization or copolymerization reactions in a multistage polymerization sequence of successive polymerization stages;
carrying out the polymerization reactions in the presence of a single site catalyst capable of forming a composition comprising
a low molecular weight component with an MFR2 of 50\u20131000 g10 min or more and a density in the range of 940 to 975 kgm3 but higher than the density of the composition, and
a high molecular weight component, said composition having a melt flow rate MFR2 of 10\u201320 g10 min and a density of 930\u2013940 kgm3, and
extruding said composition on the substrate as such or by adding 10 wt-% or less, calculated from the total weight of the coating, of high pressure PE-LD by blending into the extrusion composition or by co-extrusion;
wherein the active complex of said single site catalyst has the general formula
(X1)(X2)Hf(Cp-R1)(Cp-R2),
where X1 and X2 are thee same or different and selected from the group comprising halogen, methyl, benzyl, and hydrogen;
Hf is hafnium;
Cp is cyclopentadienyl group; and R1 and R2 are same or different and stand for linear and branched hydrocarbyl groups containing 1\u201310 carbon atoms.
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 method for transduction of hematopoietic cells by a replication-defective recombinant retrovirus vector, comprising infecting viable hematopoietic cells in culture with a replication-defective recombinant retrovirus in the presence of substantially pure fibronectin, substantially pure fibronectin fragments, or a mixture thereof, to produce transduced hematopoietic cells, said fibronectin and said fibronectin fragments containing the alternatively spliced CS-1 cell adhesion domain and the Heparin II binding domain of fibronectin.
2. The method of claim 1 which includes harvesting the transduced hematopoietic cells.
3. The method of claim 1 wherein the hematopoietic cells have a protein deficiency or abnormality and the recombinant retrovirus vector includes an exogenous gene encoding the protein.
4. The method of claim 1 wherein the hematopoietic cells have an enzyme deficiency or abnormality and the exogenous gene is a gene encoding the enzyme.
5. The method of claim 4 wherein the hematopoietic cells are human hematopoietic cells having an enzyme deficiency or abnormality and the exogenous gene is a human gene encoding the enzyme.
6. The method of claim 4 wherein the hematopoietic cells have an adenosine deaminase deficiency and the exogenous gene encodes adenosine deaminase.
7. The method of claim 5 wherein the human hematopoietic cells have an adenosine deaminase deficiency and the exogenous gene encodes adenosine deaminase.
8. The method of claim 5 wherein the cells are infected with the retrovirus in the presence of an immobilized fibronectin fragment containing an amino acid sequence which provides the cell-binding activity of the CS-1 domain and an amino acid sequence which provides the retrovirus binding activity of the Heparin-II domain.
9. The method of claim 8 wherein the fibronectin fragment is a recombinant fibronectin fragment.
10. The method of claim 9, wherein the recombinant fibronectin fragment is selected from the group consisting of H-296 and CH-296.
11. The method of claim 10, wherein the recombinant fibronectin fragment is CH-296.
12. The method of claim 9, wherein the recombinant fibronectin fragment contains the Heparin-II binding domain of fibronectin.
13. The method of claim 1, wherein the hematopoietic cells are characterized as adherent-negative, low density, mononuclear cells.
14. A method for transduction of viable mammalian cells by a replication-defective recombinant retrovirus vector, comprising infecting the cells in culture with a replication-defective recombinant retrovirus in the presence of substantially pure fibronectin, substantially pure fibronectin fragments, or a mixture thereof, without cocultivation with retrovirus producing cells to produce transduced cells.
15. The method of claim 14, wherein the infecting is in the presence of a fibronectin fragment containing the Heparin-II binding domain of fibronectin.
16. The method of claim 15, wherein said domain has an amino acid sequence represented by the formula (SEQ. ID NO. 1):
Ala Ile Pro Ala Pro Thr Asp Leu Lys Phe Thr Gln Val Thr Pro Thr Ser Leu Ser Ala Gln Trp Thr
Pro Pro Asn Val Gln Leu Thr Gly Tyr Arg Val Arg Val Thr Pro Lys Glu Lys Thr Gly Pro Met
Lys Glu Ile Asn Leu Ala Pro Asp Ser Ser Ser Val Val Val Ser Gly Leu Met Val Ala Thr Lys Tyr
Glu Val Ser Val Tyr Ala Leu Lys Asp Thr Leu Thr Ser Arg Pro Ala Gln Gly Val Val Thr Thr
Leu Glu Asn Val Ser Pro Pro Arg Arg Ala Arg Val Thr Asp Ala Thr Glu Thr Thr Ile Thr Ile Ser
Trp Arg Thr Lys Thr Glu Thr Ile Thr Gly Phe Gln Val Asp Ala Val Pro Ala Asn Gly Gln Thr Pro
Ile Gln Arg Thr Ile Lys Pro Asp Val Arg Ser Tyr Thr Ile Thr Gly Leu Gln Pro Gly Thr Asp Tyr
Lys Ile Tyr Leu Tyr Thr Leu Asn Asp Asn Ala Arg Ser Ser Pro Val Val Ile Asp Ala Ser Thr Ala
Ile Asp Ala Pro Ser Asn Leu Arg Phe Leu Ala Thr Thr Pro Asn Ser Leu Leu Val Ser Trp Gln Pro
Pro Arg Ala Arg Ile Thr Gly Tyr Ile Ile Lys Tyr Glu Lys Pro Gly Ser Pro Pro Arg Glu Val Val
Pro Arg Pro Arg Pro Gly Val Thr Glu Ala Thr Ile Thr Gly Leu Glu Pro Gly Thr Glu Tyr Thr Ile
Tyr Val Ile Ala Leu Lys Asn Asn Gln Lys Ser Glu Pro Leu Ile Gly Arg Lys Lys Thr.
17. The method of claim 16, wherein said fibronectin fragments comprise a recombinant fibronectin fragment selected from the group consisting of CH-296 and H-296.
18. An improved method of gene transfer into mammalian cells by a replication-defective recombinant retrovirus vector, the improvement comprising conducting the gene transfer without cocultivation, with retrovirus producing cells and in the presence of substantially pure fibronectin, substantially pure fibronectin fragments, or a mixture thereof, so as to increase the frequency of the gene transfer.
19. A cellular composition comprising viable hematopoietic cells transduced by retroviral-mediated gene transfer in the absence of retroviral producer cells and in the presence of an immobilized amount of a polypeptide containing fibronectin, a fibronectin fragment, or a mixture thereof, which provides the binding activity of the Heparin-II binding domain of fibronectin and an amino acid sequence which provides the cell-binding activity of the CS-1 domain of fibronectin, said immobilized amount of polypeptide being effective to increase the frequency of transduction of the hematopoietic cells by the retrovirus vector; said composition also comprising said polypeptide.
20. The cellular composition of claim 19 which is enriched in viable hematopoietic cells wherein said viable hematopoietic cells are stem cells.
21. The cellular composition of claim 19 wherein said viable hematopoietic cells are human hematopoietic cells enriched in human hematopoietic stem cells.
22. The cellular composition of claim 21 which is a substantially homogenous population of human hematopoietic cells characterized as adherent-negative, low density, mononuclear cells.
23. The cellular composition of claim 22 which has been transduced by a recombinant retrovirus vector containing an exogenous gene to correct a protein deficiency or abnormality in the cells.
24. The cellular composition of claim 20 wherein said hematopoietic cells are obtained from umbilical cord blood.