US4478934A
Expired / Lapsed
US
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Determination of adenosine by immunoassay involving acylation of the adenosine
Assignee: Yamasa Shoyu KK
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Filing: 1982-07-09
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Granted: 1984-10-23
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Expires: 2002-07-09
Abstract
Abstract A method of quantative determination of adenosine by means, of competitive immunoassay based on a competitive antigen-antibody reaction. In the competitive antigen-antibody reaction, an antibody is used which is obtained from an animal which has been immunized by introduction thereto of an antigen which comprises a carrier protein bonded with 2'- and 3'-hydroxyls of the adenosine through dicarboxylic acid residues, and a labelled adenosine and 2',3'-diacyladenosine which has been produced by acylation of adenosine in the sample to be assayed or in a standard solution are caused to undergo competitive reaction for the antibody whereby it has been made possible to determine adenosine quantitatively in high sensitivity and in high accuracy.
Inventors
- 👤 Tomokazu Sato
- 👤 Michio Ui
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 16:44
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/536; G01N33/537; G01N33/5308; Y; Y10; Y10S; Y10S436/00; Y10S436/815
Claims Analysis
Total Claims: 7
Claim 1
Independent Claim
159 words
A method for determining quantitatively the adenosine content in a sample by immunoassay, comprising the steps of: (1) adding an acylating agent to a liquid sample and to a standard adenosine solution thereby acylating hydroxyl groups at the 2'-position and the 3'-position of each adenosine; (2) diluting each of the reaction mixtures which have undergone said acylation by addition of a buffer and then carrying out antigen-antibody reactions by mixing (a) each of these reaction mixtures with (b) a predetermined amount of labelled 2',3'-diacyladenosine and (c) a predetermined amount of anti-adenosine antibodies derived from an antigen comprising adenosine bound at the 2'- and the 3'-hydroxyl groups to a carrier protein through dicarboxylic acid residues; and (3) subsequently separating, in each of the reaction mixtures, free labelled adenosine from the labelled adenosine bound to the anti-adenosine antibody and measuring the quantity of the label in either one of the labelled adenosines to calculate the adenosine content in the sample therefrom.
Claim 2
Independent Claim
159 words
A method for determining quantitatively the adenosine content in a sample by immunoassay, comprising the steps of: (1) adding an acylating agent to a liquid sample and to a standard adenosine solution thereby acylating hydroxyl groups at the 2'-position and the 3'-position of each adenosine; (2) diluting each of the reaction mixtures which have undergone said acylation by addition of a buffer and then carrying out antigen-antibody reactions by mixing (a) each of these reaction mixtures with (b) a predetermined amount of labelled 2',3'-diacyladenosine and (c) a predetermined amount of anti-adenosine antibodies derived from an antigen comprising adenosine bound at the 2'- and the 3'-hydroxyl groups to a carrier protein through dicarboxylic acid residues; and (3) subsequently separating, in each of the reaction mixtures, free labelled adenosine from the labelled adenosine bound to the anti-adenosine antibody and measuring the quantity of the label in either one of the labelled adenosines to calculate the adenosine content in the sample therefrom.
Claim 3
Dependent Claim
32 words
A method according to claim 1, wherein the dicarboxylic acid residue which binds the adenosine to the carrier protein in the antigen for obtaining the anti-adenosine antibody is a succinic acid residue.
Claim 4
Dependent Claim
18 words
A method according to claim 1, wherein the acylating agent comprises an acid anhydride and an organic text-amine.
Claim 5
Dependent Claim
13 words
A method according to claim 3, wherein the acid anhydride is succinic anhydride.
Claim 6
Dependent Claim
14 words
A method according to claim 1, wherein the labelled 2',3'-diacyladenosine is a labelled 2',3'-disuccinyladenosine.
Claim 7
Dependent Claim
20 words
A method according to claim 1, wherein the label is a radioactive element, enzyme, fluorescent substance or chemical luminescent substance.
References Cited (Backward Citations)
No citation records indexed.
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1987-10-23 | 1988-04-23 | 1988-10-23 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-10-23 | 1992-04-23 | 1992-10-23 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-10-23 | 1996-04-23 | 1996-10-23 | $7700.00 | Window Closed / Lapsed |