US4436825A
Expired / Lapsed
US
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Process and reagents for antibody detection involving erythrocyte agglutination
Assignee: Montefiore Hospital and Medical Center Inc
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Filing: 1982-05-21
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Granted: 1984-03-13
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Expires: 2002-05-21
Abstract
Abstract A process for the detection of antibodies in a test sample is described. This process involves: (a) preparing an essentially isotonic and low ionic strength suspension comprising said sample and erythrocytes in net negatively-charged form; (b) maintaining said suspension for at least 30 seconds; (c) combining said suspension with an amount of a solution of polymer effective for agglutination of said erythrocytes; (d) separating the resultant agglutinates of polymer and erythrocytes from supernatant of said suspension; (e) dispersing said agglutinates in a hypertonic salt solution having an essentially neutral pH; and (f) monitoring the dispersed agglutinates for dissociation of erythrocytes. This process permits rapid detection of antibodies with great ease. Their presence is indicated by persistence of the agglutinates in the dispersion. The antigen which reacts with antibody may be native to the erythrocytes, or may be unrelated to erythrocytes but artificially coupled to those cells. Quantitative analysis is likewise possible because their dissociation is inversely related to antibody concentration.
Inventors
- 👤 Parviz Lalezari
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 21:31
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/554; G01N33/555; Y; Y10; Y10T; Y10T436/00; Y10T436/10; Y10T436/107497
Claims Analysis
Total Claims: 49
Claim 1
Independent Claim
109 words
A process for the detection of antibodies in a test sample comprising: (a) preparing an essentially isotonic and low ionic strength suspension comprising said sample, a sensitization effective amount of ethylenediamine tetra-acetate, and erythrocytes in net negatively-charged form; (b) maintaining said suspension for at least 30 seconds; (c) combining said suspension with an amount of a solution of polymer effective for agglutination of said erythrocytes; (d) separating the resultant agglutinates of polymer and erythrocytes from supernatant of said suspension; (d) dispersing said agglutinates without bound antibody in a hypertonic salt solution having an essentially neutral pH; and (f) monitoring the resuspended agglutinates for the presence or absence of antibody.
Claim 2
Dependent Claim
109 words
A process for the detection of antibodies in a test sample comprising: (a) preparing an essentially isotonic and low ionic strength suspension comprising said sample, a sensitization effective amount of ethylenediamine tetra-acetate, and erythrocytes in net negatively-charged form; (b) maintaining said suspension for at least 30 seconds; (c) combining said suspension with an amount of a solution of polymer effective for agglutination of said erythrocytes; (d) separating the resultant agglutinates of polymer and erythrocytes from supernatant of said suspension; (d) dispersing said agglutinates without bound antibody in a hypertonic salt solution having an essentially neutral pH; and (f) monitoring the resuspended agglutinates for the presence or absence of antibody.
Claim 3
Dependent Claim
16 words
The process of claim 1 wherein the dispersed agglutinates without bound antibody are monitored for dissociation.
Claim 4
Dependent Claim
14 words
The process of claim 1 wherein the undispersed agglutinates with bound antibody are monitored.
Claim 5
Dependent Claim
104 words
A process for the detection of antibodies in a test sample comprising: (a) preparing an essentially isotonic and low ionic strength suspension comprising said sample and erythrocytes in net negatively-charged form; (b) maintaining said suspension for at least 30 seconds; (c) combining said suspension with an amount of a solution of polymer effective for agglutination of said erythrocytes; (d) separating the resulting agglutinates of polymer and erythrocytes from supernatant of said suspension; (e) dispersing said agglutinates without bound antibody in a hypertonic salt solution having a pH of from 6.8 to 7.5; (f) monitoring the resuspended agglutinates for the presence or absence of antibody.
Claim 6
Dependent Claim
104 words
A process for the detection of antibodies in a test sample comprising: (a) preparing an essentially isotonic and low ionic strength suspension comprising said sample and erythrocytes in net negatively-charged form; (b) maintaining said suspension for at least 30 seconds; (c) combining said suspension with an amount of a solution of polymer effective for agglutination of said erythrocytes; (d) separating the resulting agglutinates of polymer and erythrocytes from supernatant of said suspension; (e) dispersing said agglutinates without bound antibody in a hypertonic salt solution having a pH of from 6.8 to 7.5; (f) monitoring the resuspended agglutinates for the presence or absence of antibody.
Claim 7
Dependent Claim
16 words
The process of claim 4 wherein the dispersed agglutinates without bound antibody are monitored for dissociation.
Claim 8
Dependent Claim
14 words
The process of claim 4 wherein the undispersed agglutinates with bound antibody are monitored.
Claim 9
Dependent Claim
15 words
The process of claim 4 wherin the low ionic strength suspension further comprises ethylenediamine tetra-acetate.
Claim 10
Dependent Claim
37 words
The process of claim 4 wherein the low ionic strength suspension further comprises a sensitization effective amount of ethylenediamine tetra-acetate having a pH of from about 6.0 to 6.6, and from 3.3 to 6.7% sugar by weight.
Claim 11
Dependent Claim
42 words
The process of claim 4 wherein the hypertonic salt solution has a pH of from 6.8 to 7.5 and further comprises an aqueous solution of from about 0.1 to 0.3 molar citrate salt and from about 1.5 to 5% sugar by weight.
Claim 12
Dependent Claim
26 words
The process of claim 1, 2, 3, 4, 5, or 6, wherein the test sample is selected from the group consisting of blood and blood serum.
Claim 13
Dependent Claim
22 words
The process of claim 1, 2, 3, 4, 5, or 6, wherein the suspension comprises from 0.1 to 1% erythrocytes by volume.
Claim 14
Dependent Claim
18 words
The process of claim 1, 2, 3, 4, 5, or 6, wherein the polymer is a negatively-charged polymer.
Claim 15
Dependent Claim
18 words
The process of claim 1, 2, 3, 4, 5, or 6, wherein the polymer is a positively-charged polymer.
Claim 16
Dependent Claim
13 words
The process of claim 13, wherein the polymer is a quaternary ammonium salt.
Claim 17
Dependent Claim
11 words
The process of claim 13, wherein the polymer is hexadimethrine bromide.
Claim 18
Dependent Claim
20 words
The process of claim 1, 2, or 3, wherein the suspension has a pH of between about 6.0 and 6.6.
Claim 19
Dependent Claim
24 words
The process of claim 1, 2, or 3, wherein the agglutinates are at a temperature of from about 16° to 22° C. when dispersed.
Claim 20
Dependent Claim
24 words
The process of claim 1, 2, or 3, wherein the agglutinates are at a temperature of from about 0° to 6° C. when dispersed.
Claim 21
Dependent Claim
21 words
The process of claim 4, 5, 6, or 7 wherein antiglobulin reagent is added to the agglutinates to monitor for antibody.
Claim 22
Dependent Claim
20 words
The process of claim 1, 2, 3, 4, 5, 6, or 7 wherein the antigen is native to the erythrocytes.
Claim 23
Dependent Claim
21 words
The process of claim 1, 2, 3, 4, 5, 6, or 7 wherein the antigen is artificially coupled to the erythrocytes.
Claim 24
Dependent Claim
34 words
A reagent for use in the detection of antibodies comprising a medium having a sensitization effective pH, and containing a sensitization effective amount of ethylenediamine tetra-acetate and from 4 to 8% sugar by weight.
Claim 25
Dependent Claim
34 words
A reagent for use in the detection of antibodies comprising a medium having a sensitization effective pH, and containing a sensitization effective amount of ethylenediamine tetra-acetate and from 4 to 8% sugar by weight.
Claim 26
Dependent Claim
9 words
The reagent of claim 22, which additionally comprises erythrocytes.
Claim 27
Dependent Claim
15 words
The reagent of claim 22, which additionally comprises from 0.02 to 0.2% erythrocytes by volume.
Claim 28
Dependent Claim
17 words
The reagent of claim 22, wherein the total salt concentration is from about 0.005 to 0.025 molar.
Claim 29
Dependent Claim
11 words
The reagent of claim 25, wherein the sugar is a mono-saccharide.
Claim 30
Dependent Claim
9 words
The reagent of claim 26, which additionally comprises erythrocytes.
Claim 31
Dependent Claim
19 words
The reagent of claim 27, wherein said reagent is at a temperature of from about 16° to 22° C.
Claim 32
Dependent Claim
19 words
The reagent of claim 27, wherein said reagent is at a temperature of from about 0° to 6° C.
Claim 33
Dependent Claim
13 words
The reagent of claim 27, which additionally comprises from 0.02 to 0.2% erythrocytes.
Claim 34
Dependent Claim
38 words
A reagent for use in the detection of antibodies comprising a medium having a pH of from 6.8 to 7.5 and containing from 0.1 to 0.3 molar citrate salt and from about 1.5 to 5% sugar by weight.
Claim 35
Dependent Claim
38 words
A reagent for use in the detection of antibodies comprising a medium having a pH of from 6.8 to 7.5 and containing from 0.1 to 0.3 molar citrate salt and from about 1.5 to 5% sugar by weight.
Claim 36
Dependent Claim
19 words
The reagent of claim 31, wherein said reagent is at a temperature of from about 16° to 22° C.
Claim 37
Dependent Claim
18 words
The reagent of claim 31, wherein said reagent is at a temperature offrom about 0° to 6° C.
Claim 38
Dependent Claim
11 words
The reagent of claim 31, wherein the sugar is a mono-saccharide.
Claim 39
Dependent Claim
23 words
The reagent of claim 34, wherein the cation of the citrate salt is selected from the group consisting of sodium, potassium and ammonium.
Claim 40
Dependent Claim
125 words
A process for the detection of antibodies in a test sample comprising; (a) preparing an essentially isotonic and low ionic strength suspension comprising said sample and erythrocytes in net negatively-charged form; (b) maintaining said suspension for at least 30 seconds; (c) combining said suspension with an amount of a solution of polymer effective for agglutination of said erythrocytes; (d) separating the resultant agglutinates of polymer and erythrocytes from supernatant of said suspension; (e) dispersing said agglutinates in a hypertonic salt solution having an essentially neutral pH; (f) monitoring the dispersed agglutinates for dissociation of erythrocytes; and (g) retesting of dissociated erythrocytes for antibodies which are not detected by steps (a) through (f) by adding oligo- or poly-specific antiglobulin reagent to monitor for previously undetected antibodies.
Claim 41
Dependent Claim
125 words
A process for the detection of antibodies in a test sample comprising; (a) preparing an essentially isotonic and low ionic strength suspension comprising said sample and erythrocytes in net negatively-charged form; (b) maintaining said suspension for at least 30 seconds; (c) combining said suspension with an amount of a solution of polymer effective for agglutination of said erythrocytes; (d) separating the resultant agglutinates of polymer and erythrocytes from supernatant of said suspension; (e) dispersing said agglutinates in a hypertonic salt solution having an essentially neutral pH; (f) monitoring the dispersed agglutinates for dissociation of erythrocytes; and (g) retesting of dissociated erythrocytes for antibodies which are not detected by steps (a) through (f) by adding oligo- or poly-specific antiglobulin reagent to monitor for previously undetected antibodies.
Claim 42
Dependent Claim
123 words
A process for the detection of antibodies in a test sample comprising: (a) preparing an essentially isotonic and low ionic strength suspension comprising said sample, an aqueous solution of a sensitization effective amount of ethylenediamine tetra-acetate having a pH of from about 6.0 to 6.6, from 4 to 8% sugar by weight, and erythrocytes in net negatively-charged form; (b) maintaining said suspension for at least 30 seconds; (c) combining said suspension with an amount of a solution of polymer effective for agglutination of said erythrocytes; (d) separating the resultant agglutinates of polymer and erythrocytes from supernatant of said suspension; (e) dispersing said agglutinates in a hypertonic salt solution having an essentially neutral pH; and (f) monitoring the dispersed agglutinates for dissociation of erythrocytes.
Claim 43
Dependent Claim
123 words
A process for the detection of antibodies in a test sample comprising: (a) preparing an essentially isotonic and low ionic strength suspension comprising said sample, an aqueous solution of a sensitization effective amount of ethylenediamine tetra-acetate having a pH of from about 6.0 to 6.6, from 4 to 8% sugar by weight, and erythrocytes in net negatively-charged form; (b) maintaining said suspension for at least 30 seconds; (c) combining said suspension with an amount of a solution of polymer effective for agglutination of said erythrocytes; (d) separating the resultant agglutinates of polymer and erythrocytes from supernatant of said suspension; (e) dispersing said agglutinates in a hypertonic salt solution having an essentially neutral pH; and (f) monitoring the dispersed agglutinates for dissociation of erythrocytes.
Claim 44
Dependent Claim
109 words
A process for the detection of antibodies in a test sample comprising: (a) preparing an essentially isotonic and low ionic strength suspension comprising said sample, a sensitization effective amount of ethylenediamine tetra-acetate, and erythrocytes in net negatively-charged form; (b) maintaining said suspension for at least 30 seconds; (c) combining said suspension with an amount of a solution of polymer effective for agglutination of said erythrocytes; (d) separating the resultant agglutinates of polymer and erythrocytes from supernatant of said suspension; (e) dispersing said agglutinates in a hypertonic salt solution having a pH of from 6.8 to 7.5; and (f) monitoring the dispersed agglutinates for the presence or absence of antibody.
Claim 45
Dependent Claim
109 words
A process for the detection of antibodies in a test sample comprising: (a) preparing an essentially isotonic and low ionic strength suspension comprising said sample, a sensitization effective amount of ethylenediamine tetra-acetate, and erythrocytes in net negatively-charged form; (b) maintaining said suspension for at least 30 seconds; (c) combining said suspension with an amount of a solution of polymer effective for agglutination of said erythrocytes; (d) separating the resultant agglutinates of polymer and erythrocytes from supernatant of said suspension; (e) dispersing said agglutinates in a hypertonic salt solution having a pH of from 6.8 to 7.5; and (f) monitoring the dispersed agglutinates for the presence or absence of antibody.
Claim 46
Dependent Claim
125 words
A process for the detection of antibodies in a test sample comprising: (a) preparing an essentially isotonic and low ionic strength suspension comprising said sample and erythrocytes in net negatively-charged form; (b) maintaining said suspension for at least 30 seconds; (c) combining said suspension with an amount of a solution of polymer effective for agglutination of said erythrocytes; (d) separating the resultant agglutinates of polymer and erythrocytes from supernatant of said suspension; (e) dispersing said agglutinates in a hypertonic salt solution having an essentially neutral pH; (f) monitoring the resuspended agglutinates for the presence of antibody; (g) retesting of dissociated erythrocytes for antibodies which are not detected by steps (a) through (f) by adding oligo- or poly-specific antiglobulin reagent to monitor for previously undetected antibodies.
Claim 47
Dependent Claim
125 words
A process for the detection of antibodies in a test sample comprising: (a) preparing an essentially isotonic and low ionic strength suspension comprising said sample and erythrocytes in net negatively-charged form; (b) maintaining said suspension for at least 30 seconds; (c) combining said suspension with an amount of a solution of polymer effective for agglutination of said erythrocytes; (d) separating the resultant agglutinates of polymer and erythrocytes from supernatant of said suspension; (e) dispersing said agglutinates in a hypertonic salt solution having an essentially neutral pH; (f) monitoring the resuspended agglutinates for the presence of antibody; (g) retesting of dissociated erythrocytes for antibodies which are not detected by steps (a) through (f) by adding oligo- or poly-specific antiglobulin reagent to monitor for previously undetected antibodies.
Claim 48
Dependent Claim
37 words
A reagent for use in the detection of antibodies comprising a medium having a pH of from 6.0 to 6.6 and containing a sensitization effective amount of ethylenediamine tetra-acetate and from 4 to 8% sugar by weight.
Claim 49
Dependent Claim
37 words
A reagent for use in the detection of antibodies comprising a medium having a pH of from 6.0 to 6.6 and containing a sensitization effective amount of ethylenediamine tetra-acetate and from 4 to 8% sugar by weight.
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) | 1987-03-13 | 1987-09-12 | 1988-03-13 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 1991-03-13 | 1991-09-12 | 1992-03-13 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 1995-03-13 | 1995-09-12 | 1996-03-13 | $7700.00 | Window Closed / Lapsed |