US4397960A
Expired / Lapsed
US
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Immunoassays using F(AB')2 fragments
Assignee: Bayer Corp
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Filing: 1982-03-26
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Granted: 1983-08-09
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Expires: 2002-03-26
Abstract
Abstract Immunoassays for antigens or haptens are effected using, instead of an immunoglobulin antibody, the F(ab') 2 fragments thereof. In this way, interference from endogenous RF and Clq in the fluid under assay is avoided without the necessity of pre-treating the fluid to remove or inactivate the RF or Clq. The assays may be of various types including latex agglutination and competitive binding assays. A reagent for such use comprising F(ab') 2 fragments insolubilized on a water-insoluble substrate, especially a particulate substrate, is described.
Inventors
- 👤 Claude H. Moussebois
- 👤 Pierre L. Masson
- 👤 Jean-Pierre Vaerman
- 👤 Joseph Limet
- 👤 Cesar L. Cambiaso
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-27 07:54
CPC Classifications
A; A61; A61K; A61K39/00; A61K39/395; A61K39/44; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/563; G01N33/68; G01N33/6854; G01N33/6857
Claims Analysis
Total Claims: 17
Claim 1
Independent Claim
43 words
A method of immunoassay which comprises reacting an antigen in a fluid with insolubilized F(ab') 2 fragments of an immunoglobulin which is specific to said antigen, the reaction being effected in the substantial absence of the (whole) immunoglobulin and of F(c) fragments thereof.
Claim 2
Independent Claim
43 words
A method of immunoassay which comprises reacting an antigen in a fluid with insolubilized F(ab') 2 fragments of an immunoglobulin which is specific to said antigen, the reaction being effected in the substantial absence of the (whole) immunoglobulin and of F(c) fragments thereof.
Claim 3
Independent Claim
127 words
A method of assaying a fluid by agglutination for an antigen therein, which comprises the steps of: (a) mixing a sample of the fluid with F(ab') 2 fragments of an immunoglobulin which is specific to said antigen to form a reaction mixture, said fragments being bound to water-insoluble particles, wherein, to avoid interference from exogenous Clq and RF, the reaction mixture is substantially free from the whole immunoglobulin and the F(c) fragments thereof; (b) incubating the mixture to allow reaction between the said F(ab') 2 fragments and any said antigen present to cause agglutination of said particles; and (c) directly determining the extent to which agglutination of said particles has occurred in the mixture and thereby the presence and/or amount of said antigen in the fluid sample.
Claim 4
Independent Claim
127 words
A method of assaying a fluid by agglutination for an antigen therein, which comprises the steps of: (a) mixing a sample of the fluid with F(ab') 2 fragments of an immunoglobulin which is specific to said antigen to form a reaction mixture, said fragments being bound to water-insoluble particles, wherein, to avoid interference from exogenous Clq and RF, the reaction mixture is substantially free from the whole immunoglobulin and the F(c) fragments thereof; (b) incubating the mixture to allow reaction between the said F(ab') 2 fragments and any said antigen present to cause agglutination of said particles; and (c) directly determining the extent to which agglutination of said particles has occurred in the mixture and thereby the presence and/or amount of said antigen in the fluid sample.
Claim 5
Dependent Claim
12 words
A method according to claim 2, wherein said particles are latex particles.
Claim 6
Dependent Claim
22 words
A method according to claim 2, wherein the extent of agglutination is determined by selectively counting the agglutinated or unagglutinated latex particles.
Claim 7
Dependent Claim
32 words
A method according to claim 2, wherein said particles are magnetically attractable and wherein, after reaction between the antigen and the fragments, the particulate material is separated by using a magnetic field.
Claim 8
Dependent Claim
15 words
A method according to claim 2, wherein the fluid under assay is a biological fluid.
Claim 9
Dependent Claim
14 words
A method according to claim 2, wherein the fluid under assay is human serum.
Claim 10
Dependent Claim
13 words
A method according to claim 2, which is effected by continuous flow techniques.
Claim 11
Dependent Claim
14 words
A method according to claim 2, which is effected by a discrete manual technique.
Claim 12
Dependent Claim
39 words
A reagent for use in immunoassay which comprises a suspension of finely divided particulate material having bound thereto the F(ab') 2 fragments of an immunoglobulin, the suspension being substantially free from the said whole immunoglobulin and F(c) fragments thereof.
Claim 13
Dependent Claim
39 words
A reagent for use in immunoassay which comprises a suspension of finely divided particulate material having bound thereto the F(ab') 2 fragments of an immunoglobulin, the suspension being substantially free from the said whole immunoglobulin and F(c) fragments thereof.
Claim 14
Dependent Claim
16 words
A reagent according to claim 10 in which the said particulate material comprises magnetically attractable material.
Claim 15
Dependent Claim
15 words
A reagent according to claim 10 which is in the form of a latex suspension.
Claim 16
Independent Claim
121 words
A method of assaying a fluid by agglutination for an antigen therein, which comprises the steps of: (a) mixing a sample of the fluid with labelled F(ab') 2 fragments of an immunoglobulin which is specific to said antigen to form a reaction mixture, the labels being water-insoluble particles, wherein, to avoid interference from exogenous Clq and RF, the reaction is substantially free from the whole immunoglobulin and the F(c) fragments thereof; (b) incubating the mixture to allow a competition binding reaction between the F(ab') 2 fragments and any said antigen present; (c) analyzing the amount of labelled fragments which have reacted with the antigen or remain unreacted in the mixture to determine the presence and/or amount of antigen in the fluid.
Claim 17
Independent Claim
121 words
A method of assaying a fluid by agglutination for an antigen therein, which comprises the steps of: (a) mixing a sample of the fluid with labelled F(ab') 2 fragments of an immunoglobulin which is specific to said antigen to form a reaction mixture, the labels being water-insoluble particles, wherein, to avoid interference from exogenous Clq and RF, the reaction is substantially free from the whole immunoglobulin and the F(c) fragments thereof; (b) incubating the mixture to allow a competition binding reaction between the F(ab') 2 fragments and any said antigen present; (c) analyzing the amount of labelled fragments which have reacted with the antigen or remain unreacted in the mixture to determine the presence and/or amount of antigen in the fluid.
References Cited (Backward Citations)
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1986-08-08 | 1987-02-07 | 1987-08-09 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1990-08-08 | 1991-02-07 | 1991-08-09 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1994-08-08 | 1995-02-07 | 1995-08-09 | $7700.00 | Window Closed / Lapsed |