Apparatuses and methods for reducing albumin in samples
View Patent ↗The present invention is directed to apparatuses and methods for reducing the content of albumin in serum, plasma and/or blood samples. The apparatuses comprise an insoluble support with a ligand attached thereto. According to certain embodiments of the invention, the ligand is a bromosulfophthalein, Cibacron Blue or Warfarin ligand or salts or esters thereof. Also provided are methods of making the apparatus, as well as kits and albumin-depleted samples produced by the methods of the present invention.
1. A method for producing an albumin-depleted sample, comprising: providing a sample, which sample includes albumin and phosphate-buffered saline;
running the sample over an insoluble support having attached thereto a ligand comprising at least one hydroxyl group, said ligand consisting of bromosulfophthalein or a salt or ester thereof, said ligand attached to said insoluble support via an epoxy linkage; and
allowing albumin from the sample to bind to the ligand, thereby providing the albumin-depleted sample.
2. The method of claim 1 , further comprising collecting the albumin-depleted sample.
3. The method of claim 1 , further comprising:
running the albumin-depleted sample over the insoluble support one or more additional times; and
allowing albumin from the albumin-depleted sample to bind to the ligand, thereby providing a further albumin-depleted sample.
4. The method of claim 3 , further comprising collecting the further albumin-depleted sample.
5. The method of claim 1 , wherein the sample is selected from the group consisting of serum, plasma, and blood.
6. The method of claim 1 , wherein the albumin is human serum albumin.
7. A method for producing a protein-depleted sample, comprising:
providing a sample, which sample includes albumin and one or more additional proteins and phosphate-buffered saline; running the sample over an insoluble support having a ligand comprising at least one hydroxyl group, said ligand consisting of bromosulfophthalein or a salt or ester thereof, said ligand attached to said insoluble support via an epoxy linkage,
allowing albumin from the sample to bind to the ligand;
running the sample over one or more insoluble supports adapted to be capable of preferentially removing one or more non-albumin proteins from the sample; and
allowing the one or more non-albumin proteins to be removed from the sample, thereby providing a protein-depleted sample.
8. The method of claim 7 , further comprising collecting the protein-depleted sample.
9. The method of claim 7 , further comprising:
running the protein-depleted sample over one or more of the insoluble supports one or more additional times; and
allowing additional proteins from the protein-depleted sample to be removed, thereby providing a further protein-depleted sample.
10. The method of claim 7 , wherein the steps of running the sample over one or more other insoluble supports adapted to be capable of preferentially removing one or more non-albumin proteins from the sample and allowing the one or more non-albumin proteins to be removed from the sample are performed before running the sample over an insoluble support having a ligand capable of preferentially binding albumin and allowing albumin from the sample to bind to the ligand.
11. The method of claim 7 , wherein the albumin is human serum albumin.
12. The method of claim 5 , wherein said serum is present in a concentration of a 1:2 ratio of serum to phosphate-buffered saline.
13. The method of claim 7 , wherein the sample is selected from the group consisting of serum, plasma, and blood.
14. The method of claim 13 , wherein said serum is present in a concentration of a 1:2 ratio of serum to phosphate-buffered saline.