Tangential flow filtration devices and methods for leukocyte enrichment
First Claim
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1. A tangential flow filtration device for preparing a cell population enriched for leukocytes, comprising:
- a remover unit having a cross-flow chamber, a filtrate chamber and a filter disposed therebetween the filter having a retentate surface and a filtrate surface, the filter in fluid communication with the cross-flow chamber and the filtrate chamber;
the cross-flow chamber having an inlet and an outlet, the inlet disposed to introduce the cell population comprising a sample of blood constituents comprising leukocytes into the cross-flow chamber and parallel to the retentate surface of the filter; and
the outlet centrally disposed in a portion of the cross-flow chamber opposite the filter retentate surface;
the filter having an average pore size ranging from about 1 to about 10 microns;
such that flow of the sample of blood constituents across the filter enriches the cell population comprising the sample of blood constituents for leukocytes.
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Abstract
The present invention provides tangential flow filtration devices and methods for enriching a heterogeneous mixture of blood constituents for leukocytes by removal of non-leukocyte blood constituents. In one particular embodiment the device can provide a composition enriched in monocytes. One embodiment includes a remover unit (1) having a crossflow chamber (3) separated by a microporous filter (5) from a filtrate chamber (4), the remover unit (1) also having a tangential flow inlet (6), a fluid outlet (7) for a fluid enriched in leukocytes and a filtrate outlet (8).
63 Citations
45 Claims
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1. A tangential flow filtration device for preparing a cell population enriched for leukocytes, comprising:
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a remover unit having a cross-flow chamber, a filtrate chamber and a filter disposed therebetween the filter having a retentate surface and a filtrate surface, the filter in fluid communication with the cross-flow chamber and the filtrate chamber; the cross-flow chamber having an inlet and an outlet, the inlet disposed to introduce the cell population comprising a sample of blood constituents comprising leukocytes into the cross-flow chamber and parallel to the retentate surface of the filter; and
the outlet centrally disposed in a portion of the cross-flow chamber opposite the filter retentate surface;the filter having an average pore size ranging from about 1 to about 10 microns;
such that flow of the sample of blood constituents across the filter enriches the cell population comprising the sample of blood constituents for leukocytes. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22)
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23. A tangential flow device for enriching a cell population comprising a sample of blood constituents for leukocytes, comprising:
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a remover unit comprising a cross-flow chamber (3) and a filtrate chamber (4) separated by a filter (5), wherein the cross-flow chamber (3) has an inlet (6) and an outlet (7), the outlet centrally disposed in an upper portion of the chamber, and wherein the inlet is disposed above the filter and introduces fluid into the cross-flow chamber substantially parallel to the filter; a means for providing a predetermined input rate (14) of the cell population comprising the sample through the cross-flow chamber inlet; and a means for reducing a filtration rate (15) through the filter;
wherein the filter has a pore size of about 3 microns to about 7 microns; and
whereby the cell population comprising the sample is enriched for leukocytes in a retentate in the cross-flow chamber.
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24. A tangential flow device for enriching cell population comprising a sample comprising blood constituents for leukocytes, comprising:
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a remover unit (1) having a cross-flow chamber (3) and a filtrate chamber (4) separated by a filter (5), the cross-flow chamber having an inlet (6) and an outlet (7), the outlet disposed above the inlet and centrally disposed in an upper portion of the chamber, and wherein the filter is disposed below and substantially parallel to the cross-flow chamber inlet; means for providing a predetermined input rate (14) of the sample through the cell population comprising the sample through the cross-flow chamber inlet; means for providing a predetermined filtration rate (15) of the fluid through the filter, wherein the predetermined filtration rate is about one-fifth to about one one-hundredth of the predetermined input rate; and means for providing a predetermined concentration of blood cells in the sample, wherein the predetermined concentration of blood cells is about 107 to about 1010 cells per milliliter; wherein the filter has pores having a pore size of about 3 microns to about 7 microns; and whereby the sample is enriched for leukocytes in a retentate in the cross-flow chamber.
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25. A method for separating leukocytes from a cell population comprising a sample of blood constituents from a subject wherein the sample comprises leukocytes, the method comprising:
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(1) introducing the sample into a remover unit wherein the remover unit comprises a cross-flow chamber, a filtrate chamber and a filter disposed therebetween, the filter having a retentate surface and a filtrate surface, the filter in fluid communication with the cross-flow chamber and the filtrate chamber;
the cross-flow chamber having an inlet and an outlet, the inlet disposed to introduce the cell population comprising the sample of blood constituents comprising the leukocytes from a source into the cross-flow chamber and parallel to the retentate surface of the filter, and the outlet centrally disposed in a portion of the cross-flow chamber opposite the filter retentate surface;(2) subjecting the sample to cross-flow substantially parallel to a filter having a pore size of about 1 to about 10 microns; (3) subjecting the fluid to filtration through the filter; and (4) selectively removing non-leukocyte blood constituents from the sample through the filter and removing the leukocytes through the centrally disposed outlet to form a cell population enriched for leukocytes. - View Dependent Claims (26, 27, 28, 29, 30, 31, 32, 33, 34)
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35. A method for enriching a cell population comprising a sample of blood constituents for leukocytes, comprising:
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(1) introducing the cell population comprising the sample into a tangential flow filtration (TFF) unit, the TFF unit comprising a cross-flow chamber, a filtrate chamber, and a filter in fluid communication with the cross-flow chamber and the filtrate chamber, the filter having a retentate surface and a filtrate surface, the cross-flow chamber having an inlet and an outlet, the inlet disposed to introduce the cell population comprising the sample of blood constituents comprising leukocytes from a source into the cross-flow chamber and parallel to the retentate surface of the filter, and the outlet centrally disposed in a portion of the cross-flow chamber opposite the filter retentate surface;
the filter having a pore size of about 1 to about 10 microns;(2) recirculating the sample through the TFF unit at a predetermined input rate and a predetermined filtration rate, the predetermined input rate at least five times the predetermined filtration rate;
wherein the predetermined filtration rate is less than the unopposed filtration rate for the filter; and(3) isolating a cell population enriched for leukocytes through the outlet centrally disposed in the portion of the cross-flow chamber opposite the filter retentate surface. - View Dependent Claims (36, 37, 38, 39, 40, 41, 42, 43, 44, 45)
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Specification