Protease inhibitor sample collection system
A collection container and a method for collecting a biological sample, particularly whole blood, includes at least one stabilizing agent in an amount effective to stabilize and inhibit protein degradation and/or fragmentation. The stabilizing agent is able to stabilize proteases in the biological sample, particularly at the point of collection, by inhibiting protein degradation and/or fragmentation in the sample when the sample is stored. The stabilizing agent includes one or more protease inhibitors.
1. A sterile and evacuated tube for collecting and stabilizing whole blood or a component thereof, comprising
a first end and a second end and at least one interior wall defining a reservoir portion for receiving the blood, wherein said reservoir comprises at least one antioxidant and a protein stabilization agent comprising at least two lyophilized protease inhibitors, wherein a first inhibitor is an inhibitor of a serine protease and a second inhibitor is an inhibitor of another class of protease; and
a closure pierceable by a needle for supplying blood to said reservoir.
2. The tube of claim 1 , wherein said interior wall comprises plastic or glass.
3. The tube of claim 1 , wherein said reservoir comprises more than two lyophilized protease inhibitors.
4. The tube of claim 1 , wherein said lyophilized protease inhibitors comprise at least two inhibitors of a serine protease and at least one inhibitor of a cysteine protease.
5. The tube of claim 4 , wherein said inhibitor of a serine protease is selected from the group consisting of AEBSF, aprotinin, antithrombin, elastatinal, leupeptin, APMSF, DFP and trypsin, and mixtures thereof.
6. The tube of claim 4 , wherein said inhibitor of a serine protease comprises AEBSF, aprotinin and leupeptin.
7. The tube of claim 4 , wherein said inhibitor of a cysteine protease is E-64.
8. The tube of claim 1 , wherein said lyophilized protease inhibitors comprise at least one inhibitor of a serine protease, at least one inhibitor of a cysteine protease, and an inhibitor of a cysteine/serine protease.
9. The tube of claim 8 , wherein said inhibitor of a cysteine/serine protease is selected from the group consisting of leupeptin, TPCK, TLCK-HCl (L-1-chloro-3-[4-tosylamidol-7-amino-2-heptanone-HCl), and antipain-HCl, and mixtures thereof.
10. The tube of claim 1 , wherein said lyophilized protease inhibitors comprise at least one inhibitor of a serine protease, at least one inhibitor of a cysteine protease, and at least one of an inhibitor of a cysteine/serine protease, an inhibitor of an aspartic protease, an inhibitor of a metalloprotease, and an inhibitor of an exopeptidase.
11. The tube of claim 10 , wherein said inhibitor of an aspartic protease is selected from the group consisting of calpain, N-acetyl-leu-leu-norleucinal, N-acetyl-leu-leu-methioninal, pepstatin and VdLPFFVdL, and mixtures thereof.
12. The tube of claim 10 , wherein said inhibitor of a metalloprotease is selected from the group consisting of EDTA, 1,10-phenanthroline, phosphoramodon and mixtures thereof.
13. The tube of claim 10 , wherein said inhibitor of an exopeptidase is selected from the group consisting of amastatin, bestatin, diprotin A and diprotin B, and mixtures thereof.
14. The tube of claim 1 , wherein said at least two lyophilized protease inhibitors comprise AEBSF in a concentration of less than 1 mM at whole blood volume of the collected blood or component thereof.
15. The tube of claim 1 , further comprising an anticoagulant.
16. The tube of claim 15 , wherein said anticoagulant is spray-dried onto at least a portion of said interior wall of said tube.
17. The tube of claim 15 , wherein said anticoagulant comprises EDTA or a salt thereof.
18. The tube of claim 15 , wherein said anticoagulant comprises heparin.
19. The tube of claim 1 , further comprising collected blood.
20. The tube of claim 1 , further comprising a component of whole blood.
21. The tube of claim 20 , wherein said component of whole blood comprises serum or plasma.
22. A kit for collecting and storing a biological sample, comprising:
a primary sterile and evacuated collection tube, wherein said primary tube contains a first protein stabilization agent comprising at least one antioxidant and at least two protease inhibitors, wherein a first inhibitor is an inhibitor of a serine protease and a second inhibitor is an inhibitor of another class of protease and a closure pierceable by a needle; and
a secondary tube, wherein said secondary tube contains a second protein stabilizing agent.
23. The kit of claim 22 , further comprising a tube-to-tube transfer device.
24. The kit of claim 23 , wherein said secondary tube is maintained at a pressure to draw a sample from said primary tube through said tube-to-tube transfer device and into said secondary tube.
25. The kit of claim 22 , wherein said secondary tube is sterile.
26. The kit of claim 22 , wherein said secondary tube is evacuated.