Protease inhibitor sample collection system
A collection container and a method for collecting a biological sample, particularly whole blood, that 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 a separating element 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 piercable by a needle for supplying blood to said reservoir.
2. The tube of claim 1 , wherein the 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 , or claim 3 wherein the 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 1 , or claim 3 wherein the 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.
6. The tube of claim 1 , or claim 3 wherein the 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.
7. The tube of claim 6 , 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.
8. The tube of claim 7 , wherein said inhibitor of a serine protease comprises AEBSF, aprotinin and leupeptin.
9. The tube of claim 7 , 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.
10. The tube of claim 6 , wherein said inhibitor of a cysteine protease is E-64.
11. The tube of claim 6 , wherein said inhibitor of a cysteine/serine protease is selected from the group consisting of leupeptin, TPCK, TLCK-HCl (L-1-chloro-3-[4-tosylamido]-7-amino-2-heptanone-HCl), and antipain-HCl, and mixtures thereof.
12. The tube of claim 6 , 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.
13. The tube of claim 6 , wherein said inhibitor of a metalloprotease is selected from the group consisting of EDTA, 1,10-phenanthroline, phosphoramodon and mixtures thereof.
14. The tube of claim 6 , wherein said inhibitor of an exopeptidase is selected from the group consisting of amastatin, bestatin, diprotin A and diprotin B, and mixtures thereof.
15. The tube of claim 1 , or claim 3 wherein said separating element comprises a gel composition.
16. The tube of claim 15 , wherein said gel composition comprises a thixotropic gel.
17. The tube of claim 15 , wherein said gel composition is contained in a capsule.
18. The tube of claim 1 , or claim 3 wherein said separating element comprises a mechanical separating element.
19. The tube of claim 1 , or claim 3 further comprising an anticoagulant.
20. The tube of claim 19 , wherein said anti-coagulant is spray-dried onto at least a portion of an anterior wall of the tube.
21. The tube of claim 19 , wherein said anticoagulant comprises EDTA or a salt thereof.
22. The tube of claim 19 , wherein said anticoagulant comprises heparin.
23. The tube of claim 1 , or claim 3 further comprising collected blood.
24. The tube of claim 1 , or claim 3 further comprising a component of whole blood.
25. The tube of claim 24 , wherein said component of whole blood comprises serum or plasma.