Method of detection of analyte active forms and determination of the ability of substances to bind into analyte active sites
AssigneeUstav Organicke Chemie A Biochemie Akademie Ved Cr, V.V.I.
InventorsVaclav Navratil, Pavel Sacha, Jiri Schimer, Jan Konvalinka, Pavel Majer
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Abstract
A method for detection of active form of analytes in a sample and/or for determination of ability of tested substances to bind to the active site of these analytes has the following steps: a) analyte or group of analytes from the sample is immobilized on the surface of a solid carrier; b) analyte or group of analytes is incubated with a detection probe; c) then the solid carrier is washed to remove unbound detection probe; and subsequently, the amount of bound detection probe is determined.
Claims60
A method for detecting an analyte in a sample said method comprising the steps of:
- a) immobilizing the analyte on the surface of a solid carrier;
- b) incubating the solid carrier with
- a detection probe, wherein the detection probe consists of:
- (i) a compound capable of selectively binding to the analyte active site,
- said compound having a molecular weight of up to 2500 Da,
- (ii) an oligonucleotide tag, and
- (iii) a chemical linker covalently linking the compound and the oligonucleotide tag;
- c) washing the solid carrier to remove unbound detection probe;
- and
- d) measuring the amount of the bound detection probe directly on the solid carrier,
- thereby detecting the analyte;
- wherein the amount of the analyte in the sample is proportional to the amount of the bound detection probe.
A method for detecting an analyte in a sample, said method comprising the steps of:
- a) incubating the analyte with a detection probe, wherein the detection probe consists of:
- (i) a compound capable of selectively binding to the analyte active site, said compound having a molecular weight of up to 2500 Da,
- (ii) an oligonucleotide tag, and
- (iii) a chemical linker covalently linking the compound and the oligonucleotide tag;
- b) immobilizing the analyte on the surface of a solid carrier;
- c) washing the solid carrier to remove unbound detection probe;
- and
- d) determining the amount of the bound detection probe on the solid carrier, thereby detecting the analyte;
- wherein the amount of the analyte in the sample is proportional to the amount of the bound detection probe.
A method for measuring binding of a substance to an active site of an analyte, said method comprising the steps of:
- a) immobilizing the analyte on the surface of the solid carrier;
- b) incubating the solid carrier with the substance and a detection probe, wherein the detection probe consists of:
- (i) a compound capable of selectively binding
- to the analyte active site, said compound having a molecular weight of up to 2500 Da,
- (ii) an oligonucleotide tag, and
- (iii) a chemical linker covalently linking the compound and the oligonucleotide tag;
- c) washing the solid carrier is washed to remove unbound detection probe;
- d) measuring the amount of the bound detection probe;
- and
- e) comparing the amount of the bound detection probe with a control amount of the bound detection probe measured in the method in the absence of the substance, thereby measuring the binding of the substance to the active site of the analyte.
A method for measuring binding of a substance to an active site of an analyte, said method comprising the steps of:
- a) incubating the analyte with the substance and a detection probe, wherein the detection probe consists of:
- (i) a compound capable of selectively binding to the analyte, said compound having a molecular weight of up to 2500 Da,
- (ii) an oligonucleotide tag, and
- (iii) a chemical linker covalently linking the compound and the oligonucleotide tag;
- b) immobilizing the analyte on the surface of the solid carrier;
- c) washing the solid carrier to remove unbound detection probe;
- d) measuring the amount of the bound detection probe;
- and
- e) comparing the amount of the bound detection probe with a control amount of the bound detection probe determined in the absence of the substance,
- thereby measuring the binding of the substance to the active site of the analyte.
Dependencies are inferred from the claim text.