Title:
A METHOD FOR MEASURING BIOMOLECULES
Document Type and Number:
WIPO Patent Application WO/2000/068415
Kind Code:
A2
Abstract:
Described herein is a method for detecting molecules with biological activity and determining the identity, amount and activity of same. The method involves the use of a vessel-based system which facilitates and simplifies the measurement of the amount and activity of enzymes, enzyme inhibitors, lectins, receptors and other biologically active substances comprising the steps of carrying out a biological reaction in a first vessel and then separating a labeled reactant from a labeled product by transferring a portion of the reaction mixture to a second vessel. Specifically, the surface of the first vessel is coated with a reactant, additional components are added, thereby forming a reaction mix, and the reaction is allowed to proceed for a given period of time. The soluble portion of the reaction mix is then transferred to a second vessel and the amount of reactant liberated into the reaction mix is counted.
Inventors:
MARQUARDT RONALD R (CA)
GAN ZHIBO (CA)
GAN ZHIBO (CA)
Application Number:
PCT/CA2000/000540
Publication Date:
November 16, 2000
Filing Date:
May 10, 2000
Export Citation:
Assignee:
NORZYME INC (CA)
UNIV MANITOBA (CA)
MARQUARDT RONALD R (CA)
GAN ZHIBO (CA)
UNIV MANITOBA (CA)
MARQUARDT RONALD R (CA)
GAN ZHIBO (CA)
International Classes:
C12Q1/00; G01N33/543; (IPC1-7): C12Q1/00
Domestic Patent References:
| WO1997043438A1 | 1997-11-20 |
Foreign References:
| EP0308704A1 | 1989-03-29 | |||
| EP0296036A2 | 1988-12-21 | |||
| US5538858A | 1996-07-23 | |||
| EP0518557A2 | 1992-12-16 |
Other References:
TWINNING S S: "FLUORESCEIN ISOTHIOCYANATE-LABELED CASEIN ASSAY FOR PROTEOLYTIC ENZYMES" ANALYTICAL BIOCHEMISTRY, vol. 143, 1984, pages 30-34, XP000961435 cited in the application
JONES LAURIE J; UPSON ROSALYN H; HAUGLAND ROSARIA P; PANCHUK-VOLOSHINA NATALIYA; ZHOU MINGJIE; HAUGLAND RICHARD P: "Quenched BODIPY dye-labeled casein substrates for the assay of protease activity by direct fluorescence measurement" ANALYTICAL BIOCHEMISTRY, vol. 251, 1997, pages 144-152, XP002152059 cited in the application
JONES LAURIE J; UPSON ROSALYN H; HAUGLAND ROSARIA P; PANCHUK-VOLOSHINA NATALIYA; ZHOU MINGJIE; HAUGLAND RICHARD P: "Quenched BODIPY dye-labeled casein substrates for the assay of protease activity by direct fluorescence measurement" ANALYTICAL BIOCHEMISTRY, vol. 251, 1997, pages 144-152, XP002152059 cited in the application
Attorney, Agent or Firm:
Collard, Christine J. (60 Queen Street Ottawa Ontario, K1P 5Y7, CA)
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Claims:
CLAIMS
| 1. | A method for measuring the activity or concentration of a bioactive molecule comprising: coating a reaction vessel with a reactant, said reactant being capable of interacting with a bioactive molecule having a biological activity; adding a sample to the reaction vessel, said sample comprising a known quantity of a detectable label and the bioactive molecule having the biological activity; incubating the reaction vessel under conditions wherein the reactant and the sample interact; removing a quantity of detectable label from the sample by binding detectable label to the reactant coated on the reaction vessel; transferring a soluble portion of the sample from the reaction vessel to a counting vessel; and measuring the quantity of detectable label in the counting vessel. |
| 2. | The method according to claim 1 wherein the detectable label is selected from the group consisting of : enzyme label; colorimetric label; radioactive label; luminescent label and fluorescent label. |
| 3. | The method according to claim 1 wherein the sample is a biological sample. |
| 4. | The method according to claim 1 wherein the biological activity is an enzymatic activity. |
| 5. | The method according to claim 1 wherein the biological activity is a binding affinity. |
| 6. | The method according to claim 1 wherein the sample includes an inhibitor of the biological activity of the bioactive molecule. |
| 7. | The method according to claim 1 wherein the sample includes a competitor of the biological activity of the bioactive molecule. |
| 8. | The method according to claim 1 wherein the bioactive molecule is selected from the group consisting of: an enzymatic product; an enzyme; a substrate; a lectin; a lectinbinding ligand; a receptor; an inhibitor; a receptor binding ligand; an antigen; and an antibody. |
| 9. | The method according to claim 1 wherein the compound is selected from the group consisting of: an enzymatic product; an enzyme; a lectin; a lectinbinding ligand; a substrate; a receptor; an inhibitor; a receptor binding ligand; an antigen; and an antibody. |
| 10. | A method for measuring the activity or concentration of a bioactive molecule comprising: coating a reaction vessel with a reactant, said reactant being capable of interacting with a bioactive molecule, said reactant including a detectable label; adding to the reaction vessel a sample, said sample including the bioactive molecule having a biological activity; releasing a quantity of detectable label from the reactant by incubating the reaction vessel under conditions such that the reactant and the detectable label contact the bioactive molecule and interact with the bioactive molecule; transferring a soluble portion of the sample containing released label from the reaction vessel to a counting vessel; and measuring the quantity of detectable label in the counting vessel. |
| 11. | The method according to claim 10 wherein the detectable label is selected from the group consisting of: enzyme label; colorimetric label; radioactive label; luminescent label; and fluorescent label. |
| 12. | The method according to claim 10 wherein the sample is a biological sample. |
| 13. | The method according to claim 10 wherein the biological activity is an enzymatic activity. |
| 14. | The method according to claim 10 wherein the biological activity is a binding affinity. |
| 15. | The method according to claim 10 wherein the sample includes an inhibitor of the biological activity of the bioactive molecule. |
| 16. | The method according to claim 10 wherein the sample includes a competitor of the biological activity of the bioactive molecule. |
| 17. | The method according to claim 10 wherein the bioactive molecule is selected from the group consisting of: an enzymatic product; an enzyme; a substrate; a receptor; a receptor ligand; an antigen; a lectin; a lectinbinding ligand; a ligand; and an antibody. |
Description:
INTERNATIONALSEARCHREPORT lnternw I ApplicationNo PCT/CA00/00540 C.(Coritinuation) DOCUMENTS CONSIDEREDTOBERELEVANT Category* Citationofdocument,withindication,whereappropriate,oftherele
vantpassagesRelevanttodaimNo. AEP 0518557A(LILLYCOELI) 16December1992(1992-12-16) example1 ATWINNINGSS:"FLUORESCEIN ISOTHIOCYANATE-LABELEDCASEINASSAYFOR PROTEOLYTICENZYMES" ANALYTICALBIOCHEMISTRY, vol.143,1984,pages30-34,XP000961435 citedintheapplication thewholedocument AJONESLAURIEJ;UPSONROSALYNH;HAUGLAND ROSARIAP;PANCHUK-VOLOSHINANATALIYA; ZHOUMINGJIE;HAUGLANDRICHARDP: "QuenchedBODIPYdye-labeledcasein substratesfortheassayofprotease activitybydirectfluorescence measurement" ANALYTICALBIOCHEMISTRY, vol.251,1997,pages144-152,XP002152059 citedintheapplication abstract 1 INTERNATIONALSEARCHREPORT IMernat AppUcatlonNo tmufmatton on patent tami) y members,, PatentdocumentPublicabon Patent family Publicaton citedinsearchreportdatemember (s) date EP0308704A29-03-1989JP1063863A09-03-1989 JP1063864A09-03-1989 EP0296036A21-12-1988JP1221665A05-09-1989 WO9743438A20-11-1997AU3122497A05-12-1997 CA2254887A20-11-1997 EP0912757A06-05-1999 US5538858A23-07-1996NONE EP0518557A16-12-1992US5235039A10-08-1993 AT141649T15-09-1996 CA2070719A11-12-1992 DE69212911D26-09-1996 DK518557T09-09-1996 JP5194591A03-08-1993