Methods and devices for the enrichment of immunoglobulin from blood
A method for extracting at least 55% of immunoglobulin such as IgG from a biological fluid. The biological fluid containing immunoglobulin is contacted with a solid support covalently bonded to a ligand that specifically binds to immunoglobulin under conditions sufficient for non-covalent binding of immunoglobulin to the ligand. The solid support is contacted with an elution solution under conditions whereby the non-covalently bound immunoglobulin is released from the ligand and into the elution solution, wherein at least 55% of the IgG present in the biological fluid is extracted into the elution solution. The method, in some embodiments, provides a method for enriching immunoglobulin from a biological fluid comprising obtaining an initial biological fluid suspected of containing immunoglobulin and removing non-immunoglobulin components naturally occurring in the initial biological fluid to obtain a non-immunoglobulin component-reduced biological fluid.
1 . A method for collecting immunoglobulin from whole blood from a donor, the method comprising:
providing a set including
a separation container having an inlet and an outlet, and comprising a solid support covalently bonded to a ligand that specifically binds to immunoglobulin, wherein the separation container is configured to support the plasma product passing through the separation container at a flow rate of between about 20 ml/minute to about 120 ml/minute, and to support the plasma product passing through the separation container with a backpressure of between about 10 mm Hg to about 100 mmHg;
a blood cell collection bag having an inlet and an outlet;
a centrifuge bowl coupled to:
(a) the inlet of the separation container, the centrifuge bowl being configured to separate a plasma product from the whole blood and direct the plasma product to the inlet of the separation container, and
(b) the inlet of the blood cell collection bag; and
a collection container coupled to the outlet of the separation container, and adapted for collecting purified immunoglobulin product, wherein the separation container is in-line with the centrifuge bowl and the collection container so as to form a closed system to prevent the loss of sterility within the set, the separation container, the blood cell collection bag, the centrifuge bowl, and the collection container remain coupled as parts of the set while the plasma product is being separated from the whole blood and while the immunoglobulin is binding to the ligand in the separation container;
withdrawing the whole blood from a donor;
supplementing the whole blood with an anticoagulant;
introducing the withdrawn whole blood into the centrifuge bowl, spinning the centrifuge bowl to separate the plasma product from other blood components of the whole blood, directing the plasma product to the separation container, and directing the other blood components to the blood cell collection bag;
after the plasma product has been directed to the separation container, contacting the plasma product with the solid support covalently bonded to the ligand that specifically binds to immunoglobulin in the separation container under conditions sufficient for non-covalent binding of the immunoglobulin to the ligand;
after the non-covalent binding of the immunoglobulin to the ligand, contacting the solid support in the container with an elution solution under conditions whereby the non-covalently bound immunoglobulin is released from the ligand and into the elution solution to obtain elution solution comprising the immunoglobulin;
collecting the eluted immunoglobulin in the collection container; and
returning one or more of the other blood components from the blood cell collection bag to the donor while continuing to withdraw additional whole blood from the donor, such that the method can be performed chair-side by the donor.
2 . A method for collecting an immunoglobulin-enriched plasma product from whole blood from a donor, the method comprising:
providing a set including
a separation container having an inlet and an outlet, and comprising a solid support covalently bonded to a ligand that specifically binds to a non-immunoglobulin component in the whole blood, wherein the separation container is configured to support the plasma product passing through the separation container at a flow rate of between about 20 ml/minute to about 120 ml/minute, and to support the plasma product passing through the separation container with a backpressure of between about 10 mm Hg to about 100 mmHg;
a blood cell collection bag having an inlet and an outlet;
a centrifuge bowl coupled to:
(a) the inlet of the separation container, the centrifuge bowl being configured to separate a plasma product from the whole blood and direct the plasma product to the inlet of the separation container, and
(b) the inlet of the blood cell collection bag; and
a collection container coupled to the outlet of the separation container, and adapted for collecting immunoglobulin-enriched plasma product, wherein the separation container is in-line with the centrifuge bowl and the collection container so as to form a closed system to prevent the loss of sterility within the set, the separation container, the blood cell collection bag, the centrifuge bowl, and the collection container remain coupled as parts of the set while the plasma product is being separated from the whole blood and while the non-immunoglobulin component is binding to the ligand in the separation container;
withdrawing the whole blood from a donor;
supplementing the whole blood with an anticoagulant;
introducing the whole blood into the centrifuge bowl, spinning the centrifuge bowl to separate the plasma product from other blood components of the whole blood, directing the plasma product to the separation container, and directing the other blood components to the blood cell collection bag;
after the plasma product has been directed to the separation container, contacting the plasma product with the solid support in the separation container under conditions sufficient for non-covalent binding of the non-immunoglobulin component to the ligand;
after the non-covalent binding of the non-immunoglobulin component to the ligand, collecting the immunoglobulin-enriched plasma product in the collection container, depleted of the non-immunoglobulin component from the separation container; and
returning one or more of the other blood components from the blood cell collection bag to the donor while continuing to withdraw additional whole blood from the donor, such that the method can be performed chair-side by the donor.
3 . The method of claim 1 , wherein the method is a partially or fully automated method.
4 . The method of claim 1 , wherein the solid support are beads.
5 . The method of claim 1 , wherein the ligand is a protein from a bacteria family selected from the group consisting of Staphylococcus or Streptococcus , or a derivative thereof.
6 . The method of claim 1 , wherein the plasma product is platelet-enriched plasma or plasma.
7 . The method of claim 1 , wherein the separation container is a column or bag.
8 . The method of claim 1 , wherein the executed method further comprises adding a neutralizing buffer to the elution solution comprising immunoglobulin to result in a final solution comprising immunoglobulin having a pH of between about 7.0 and about 8.0.
9 . The method of claim 2 , wherein the solid support are beads.
10 . The method of claim 2 , wherein the plasma product is platelet-enriched plasma or plasma.
11 . The method of claim 2 , wherein the separation container is a column or bag.