US4474877A
Expired / Lapsed
US
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Process for detection and measurement of viral specific immunoglobulins and article of manufacture therefor
Assignee: Lundquist Institute for Biomedical Innovation at Harbor-UCLA Medical Center
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Filing: 1980-09-19
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Granted: 1984-10-02
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Expires: 2000-09-19
Abstract
Abstract This invention relates a simplified and reliable process for the detection and measurement of viral specific immunoglobulins such as IgG, IgM and IgE wherein relatively large-diameter plastic beads, e.g., 1/4'', diameter polystyrene balls with a specular finish, are used as the solid matrix upon which a high concentration of standard cells are first grown or cultured. The cells are then infected with a particular virus to express a high concentration of unpurified viral antigen(s), and are then fixed to preserve the viral antigen(s) concentration. The resulting fixed plastic beads (PB) are washed (to remove fixing agent) and then employed as the basis of the detection of specific immunoglobulins in the patient's serum. The detection process involves simply incubating a measured dilution of patient serum with each PB whereby the various specific immunoglobulins in the patient serum will attach to the viral antigen(s) on the PB. The specific immunoglobulins (e.g. IgG) desired to be measured is detected by the addition, to the PB container, of the anti-human specific immunoglobulin (e.g., anti-human IgG) in tagged format, e.g., in radioactive form. The tagged anti-human specific immunoglobulin binds to the specific immunoglobulin and by measuring the amount of the tagged anti-human specific anti-globulin, one obtains a direct measurement of the amount of the desired specific immunoglobulin present in the patient serum. The process techniques offer the advantages of easy preparation, storage and handling of the basic unit (PB) used for measurement, and offers increased sensitivity and substantial savings in assay time over prior art methods.
Inventors
- 👤 David T. Imagawa
- 👤 Hong D. Bui
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-31 07:30
CPC Classifications
C; C12; C12Q; C12Q1/00; C12Q1/02; C12Q1/70; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/544; G01N33/545; G01N33/554; G01N33/569; G01N33/56983; Y; Y10; Y10S; Y10S435/00; Y10S435/81; Y10S435/975; Y10S436/00; Y10S436/807; Y10S436/808
Claims Analysis
Total Claims: 10
Claim 1
Independent Claim
255 words
A process for detecting viral specific immunoglobulins in a fluid, which comprises: growing cells on a polymeric solid phase carrier of a size and weight sufficiently great to be separable from an aqueous medium without centrifugation; infecting the cells, grown on said polymeric solid phase carrier, with a particular virus of a single class to produce unpurified viral antigen (s) therein; immobilizing the said infected cells with a fixing agent of the type which destroys the viral infectiousness of said infected cells, but which preserves essentially completely, the concentration of said unpurified viral antigen(s) at the level at which it was initially produced; incubating the said polymeric solid phase carrier with fluid to be measured selected from the group consisting of patient serum, cerebrospinal fluid (CSF), negative control serum and negative control CSF at approximately room temperature and for a predetermined period of time to thereby bind different types of specific immunoglobulins in said fluid to available binding sites on the said unpurified viral antigen(s); again incubating said polymeric solid phase carrier with tagged anti-human specific immunoglobulin of the type wherein the specific immunoglobulin chosen is that desired to be measured, at approximately room temperature and for a predetermined period of time whereby a precise amount of said tagged anti-human specific immunoglobulin is bound to the specific immunoglobulin desired to be measured; removing excess, unbound tagged anti-human specific immunoglobulin; and measuring the amount of bound tagged anti-human specific immunoglobulin to thereby quantify the amount of specific immunoglobulin, desired to be measured, in the patient serum.
Claim 2
Dependent Claim
255 words
A process for detecting viral specific immunoglobulins in a fluid, which comprises: growing cells on a polymeric solid phase carrier of a size and weight sufficiently great to be separable from an aqueous medium without centrifugation; infecting the cells, grown on said polymeric solid phase carrier, with a particular virus of a single class to produce unpurified viral antigen (s) therein; immobilizing the said infected cells with a fixing agent of the type which destroys the viral infectiousness of said infected cells, but which preserves essentially completely, the concentration of said unpurified viral antigen(s) at the level at which it was initially produced; incubating the said polymeric solid phase carrier with fluid to be measured selected from the group consisting of patient serum, cerebrospinal fluid (CSF), negative control serum and negative control CSF at approximately room temperature and for a predetermined period of time to thereby bind different types of specific immunoglobulins in said fluid to available binding sites on the said unpurified viral antigen(s); again incubating said polymeric solid phase carrier with tagged anti-human specific immunoglobulin of the type wherein the specific immunoglobulin chosen is that desired to be measured, at approximately room temperature and for a predetermined period of time whereby a precise amount of said tagged anti-human specific immunoglobulin is bound to the specific immunoglobulin desired to be measured; removing excess, unbound tagged anti-human specific immunoglobulin; and measuring the amount of bound tagged anti-human specific immunoglobulin to thereby quantify the amount of specific immunoglobulin, desired to be measured, in the patient serum.
Claim 3
Dependent Claim
25 words
The process of claim 1 which includes removing said fixing agent from said solid phase carrier prior to incubation with the fluid to be measured.
Claim 4
Dependent Claim
28 words
The process of claim 1 which includes blocking free binding sites on said solid phase carriers from binding to tagged anti-human specific immunoglobulin or other protein specific immunoglobulin.
Claim 5
Dependent Claim
21 words
The process of claim 1 wherein said solid phase carrier is a plastic ball having a diameter of between 4-8 mm.
Claim 6
Dependent Claim
20 words
The process of claim 4 wherein said plastic ball is made of polystyrene and is provided with a specular surface.
Claim 7
Dependent Claim
26 words
The process of claim 1 wherein the viral specific immunoglobulins to be measured are selected from the group consisting of IgG, IgM, IgE, IgA and IgD.
Claim 8
Dependent Claim
24 words
The process of claim 1 wherein said virus for infecting said cells is selected from the group consisting of measles virus and rubella virus.
Claim 9
Dependent Claim
115 words
In a kit for the assay of viral specific immunoglobulin having a plurality of testing cavities, a plurality of polymeric solid phase carriers, having a specular surface to which viral antigens are attached, having a diameter of between 4 millimeters and about 8 millimeters and having a density sufficient to be separated from a medium without centrifugation, standards, and a tagged anti-human specific immunoglobulin, the carriers, standards, and immunoglobulins being present in sufficient amounts to perform the assay, wherein the improvement comprises: the attachment of the viral antigen to said carrier by growing cells on said carrier which cells are infected with virus to produce said viral antigens which are used in an unpurified form.
Claim 10
Dependent Claim
115 words
In a kit for the assay of viral specific immunoglobulin having a plurality of testing cavities, a plurality of polymeric solid phase carriers, having a specular surface to which viral antigens are attached, having a diameter of between 4 millimeters and about 8 millimeters and having a density sufficient to be separated from a medium without centrifugation, standards, and a tagged anti-human specific immunoglobulin, the carriers, standards, and immunoglobulins being present in sufficient amounts to perform the assay, wherein the improvement comprises: the attachment of the viral antigen to said carrier by growing cells on said carrier which cells are infected with virus to produce said viral antigens which are used in an unpurified form.
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USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 1987-10-02 | 1988-04-02 | 1988-10-02 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-10-02 | 1992-04-02 | 1992-10-02 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-10-02 | 1996-04-02 | 1996-10-02 | $7700.00 | Window Closed / Lapsed |