Toxin activity assays, devices, methods and systems therefor
Embodiments of the present invention are directed toward devices, system and method for conducting toxin activity assay using sedimentation. The toxin activity assay may include generating complexes which bind to a plurality of beads in a fluid sample. The complexes may include a target toxin and a labeling agent, or may be generated due to presence of active target toxin and/or labeling agent designed to be incorporated into complexes responsive to the presence of target active toxin. The plurality of beads including the complexes may be transported through a density media, wherein the density media has a lower density than a density of the beads and higher than a density of the fluid sample, and wherein the transporting occurs, at least in part, by sedimentation. Signal may be detected from the labeling agents of the complexes.
1. A method of conducting a toxin activity assay, the method comprising:
generating a plurality of complexes on a plurality of beads by action of an active toxin in a fluid sample, individual complexes of the plurality of complexes comprising a capture agent, and a labeling agent;
transporting the plurality of beads including the complexes through a density medium, wherein the density medium has a density lower than a density of the beads and higher than a density of the fluid sample, and wherein the transporting occurs, at least in part, by sedimentation; and
detecting a signal from the labeling agents of the plurality of complexes bound to the plurality of beads.
2. The method of claim 1 , wherein said plurality of complexes further comprises a target analyte.
3. The method of claim 2 , wherein the target analyte comprises a Staphylococcal enterotoxin B.
4. The method of claim 1 , wherein the active toxin is Ricin.
5. The method of claim 1 , wherein the active toxin is a Shiga-like toxin.
6. The method of claim 1 , wherein the active toxin is a Staphylococcal enterotoxin B.
7. The method of claim 1 , wherein the beads comprise silica beads.
8. The method of claim 7 , wherein the beads are linked to a DNA or RNA fragment.
9. The method of claim 8 , wherein the DNA or RNA fragment is a sarcin-ricin loop.
10. The method of claim 7 , wherein the beads are linked to major histocompatibility complex II.
11. The method of claim 1 , wherein the labeling agent comprises Apurinic/apyrimidinic (AP) endonuclease (APE1) enzyme.
12. The method of claim 1 , wherein the labeling agent comprises a stained immortalized T-cell line.
13. The method of claim 12 , wherein the immortalized T-cell line comprises Jurkat cells.
14. The method of claim 13 , wherein the Jurkat cells are fixed.
15. The method of claim 14 , wherein the Jurkat cells are stained by acridine orange.
16. The method of claim 1 , wherein the density medium has a density between about 1.095 g/cc and about 2.05 g/cc.
17. The method of claim 1 , wherein said fluid sample comprises whole blood, wherein the density medium has a density greater than a density of red blood cells and the method further comprises:
separating the red blood cells from the plurality of beads at least in part by applying sedimentation forces to the fluid sample such that the red blood cells remain at an interface between the density medium and fluid sample.
18. A method of conducting a toxin activity assay, the method comprising:
generating a plurality of complexes on a plurality of beads by action of an active toxin in a fluid sample, individual complexes of the plurality of complexes comprising a sarcin-ricin loop capture agent, and a labeling agent;
transporting the plurality of beads including the plurality of complexes through a density medium, wherein the density medium has a density lower than a density of the beads and higher than a density of the fluid sample, and wherein the transporting occurs, at least in part, by sedimentation; and
detecting a signal from the labeling agents of the plurality of complexes bound to the plurality of beads.
19. The apparatus of claim 18 , wherein the labeling agent comprises Apurinic/apyrimidinic (AP) endonuclease (APE1) enzyme.
20. The method of claim 19 , wherein the active toxin is Ricin.
21. The method of claim 19 , wherein the active toxin is a Shiga-line toxin.
22. A method of conducting a toxin activity assay, the method comprising:
generating a plurality of complexes on a plurality of beads by action of an active toxin in a fluid sample, individual complexes of the plurality of complexes comprising major histocompatibility complex II (MHC) capture agent, and a labeling agent;
transporting the plurality of beads including the plurality of complexes through a density medium, wherein the density medium has a density lower than a density of the beads and higher than a density of the fluid sample, and wherein the transporting occurs, at least in part, by sedimentation; and
detecting a signal from the labeling agents of the plurality of complexes bound to the plurality of beads.
23. The method of claim 22 , wherein the active toxin is a Staphylococcal enterotoxin B (SEB).
24. The method of claim 23 , wherein the labeling agent includes T cell receptors (TCR).
25. The method of claim 24 , wherein the TCR include Jurkat cells.