CA2300687A1 - Novel elisa testing system - Google Patents
Novel elisa testing system Download PDFInfo
- Publication number
- CA2300687A1 CA2300687A1 CA 2300687 CA2300687A CA2300687A1 CA 2300687 A1 CA2300687 A1 CA 2300687A1 CA 2300687 CA2300687 CA 2300687 CA 2300687 A CA2300687 A CA 2300687A CA 2300687 A1 CA2300687 A1 CA 2300687A1
- Authority
- CA
- Canada
- Prior art keywords
- antibodies
- antigens
- well
- polystyrene
- enzyme
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
Abstract
Enzyme-linked immunosorbent assay (ELISA) is perhaps one of the most widespread assay methods used extensively in the field of immunology. This assay works on the basis of known amounts of antigens or antibodies adhered onto a solid surface made of polystyrene. The polystyrene-made wells are opened at only one side and each well is eight mm in diameter, ten mm in depth and holds up to a maximum of 300 µL, in volume.
Every 8 or 12 wells joins together to form a 96-well or a 48-well combination unit. The inner wall of each well is smooth with a transparent flat base. Generally, approximately 100 µL of known antigens or antibodies is added to each well occupying one-third of the well.
Known amounts of antigens or antibodies are allowed to adhere onto the polystyrene surface at a specific temperature for over 10 hours. This process is often known as "coating". Following this, protein solutions at high concentrations or solutions with concentrated proteins and glucose are allowed to further adhere onto the polystyrene wells to ensure maximum occupancy of the well surfaces. In addition, this second coating offers better protection on the activity of the antigens or antibodies. This second process is termed "blocking".
The commonly used enzyme (horseradish peroxidase) can be linked either to the antigens or the antibodies, but in either case the solid-phase reactant is unlabeled.
ELISA can be made quantitative by setting it up as a competitive inhibition assay in which free or unknown amounts of antigens (or antibodies) in samples compete with enzyme-labeled antigens (or antibodies) for binding sites on the polystyrene solid phase coated with known amounts of antigens or antibodies. Following appropriate washings, the concentration of enzyme bound to the immune complex is estimated by the addition of its substrate that changes colour when the substrate is acted on by the enzyme.
This highly-specific immunoassay has thusfar become one of the most popular immunoassays in immunology and immunology-related studies.
Every 8 or 12 wells joins together to form a 96-well or a 48-well combination unit. The inner wall of each well is smooth with a transparent flat base. Generally, approximately 100 µL of known antigens or antibodies is added to each well occupying one-third of the well.
Known amounts of antigens or antibodies are allowed to adhere onto the polystyrene surface at a specific temperature for over 10 hours. This process is often known as "coating". Following this, protein solutions at high concentrations or solutions with concentrated proteins and glucose are allowed to further adhere onto the polystyrene wells to ensure maximum occupancy of the well surfaces. In addition, this second coating offers better protection on the activity of the antigens or antibodies. This second process is termed "blocking".
The commonly used enzyme (horseradish peroxidase) can be linked either to the antigens or the antibodies, but in either case the solid-phase reactant is unlabeled.
ELISA can be made quantitative by setting it up as a competitive inhibition assay in which free or unknown amounts of antigens (or antibodies) in samples compete with enzyme-labeled antigens (or antibodies) for binding sites on the polystyrene solid phase coated with known amounts of antigens or antibodies. Following appropriate washings, the concentration of enzyme bound to the immune complex is estimated by the addition of its substrate that changes colour when the substrate is acted on by the enzyme.
This highly-specific immunoassay has thusfar become one of the most popular immunoassays in immunology and immunology-related studies.
Description
SPECIFICATION:
This patent application relates to the development of a novel ELISA operating system in contrast to the operation of a conventional ELISA system. This innovative idea arises during the process of investigation and research on the use of an immuno-solid dry reagents system with the fully automatic analytical machine.
To fully optimize the adherence capacity of the antigens or antibodies, the surface area is increased by creating grooves throughout the lower half inner walls of the polystyrene wells. This new ELISA system uses fully solidified reagents and has two distinct differences from the conventional ELISA system in terms of operation. Firstly, the reactions of both the unknown number of antigens (or antibodies) in samples and the enzyme-labeled antigens (or antibodies) are stationary using the conventional ELISA
method. In this new ELISA system, however, the reactions of both the samples and enzyme-labeled antigens (or antibodies) are carried out under shaking at a specific temperature. In addition, the shaking frequency varies with different cases or examples.
Secondly, the washing method following the reactions of both the samples and enzyme-labeled antigens (or antibodies) is also different. The conventional ELISA
system adopts a vertical flow washing mode whereas this new ELISA system uses a liquid-rotation washing method.
This patent application pertains to the use of immuno-solid dry reagents system in conjunction with the fully automatic analytical machine. The novel idea of a special glass-frosting treatment and unique washing method are both creative changes in this new ELISA testing system. These unique treatment and changes will definitely increase the overall sensitivity and specificity of a more advanced ELISA system.
This patent application relates to the development of a novel ELISA operating system in contrast to the operation of a conventional ELISA system. This innovative idea arises during the process of investigation and research on the use of an immuno-solid dry reagents system with the fully automatic analytical machine.
To fully optimize the adherence capacity of the antigens or antibodies, the surface area is increased by creating grooves throughout the lower half inner walls of the polystyrene wells. This new ELISA system uses fully solidified reagents and has two distinct differences from the conventional ELISA system in terms of operation. Firstly, the reactions of both the unknown number of antigens (or antibodies) in samples and the enzyme-labeled antigens (or antibodies) are stationary using the conventional ELISA
method. In this new ELISA system, however, the reactions of both the samples and enzyme-labeled antigens (or antibodies) are carried out under shaking at a specific temperature. In addition, the shaking frequency varies with different cases or examples.
Secondly, the washing method following the reactions of both the samples and enzyme-labeled antigens (or antibodies) is also different. The conventional ELISA
system adopts a vertical flow washing mode whereas this new ELISA system uses a liquid-rotation washing method.
This patent application pertains to the use of immuno-solid dry reagents system in conjunction with the fully automatic analytical machine. The novel idea of a special glass-frosting treatment and unique washing method are both creative changes in this new ELISA testing system. These unique treatment and changes will definitely increase the overall sensitivity and specificity of a more advanced ELISA system.
Claims
CLAIMS:
The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:
a. The creation of grooves to increase the surface area of all polystyrene adhering surfaces;
b. The entire reaction process of this new ELISA system is carried out under a shaking condition at a specific frequency;
c. This new ELISA system adopts a liquid-rotation washing method on all washable polystyrene surfaces.
The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:
a. The creation of grooves to increase the surface area of all polystyrene adhering surfaces;
b. The entire reaction process of this new ELISA system is carried out under a shaking condition at a specific frequency;
c. This new ELISA system adopts a liquid-rotation washing method on all washable polystyrene surfaces.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA 2300687 CA2300687A1 (en) | 2000-02-23 | 2000-02-23 | Novel elisa testing system |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CA 2300687 CA2300687A1 (en) | 2000-02-23 | 2000-02-23 | Novel elisa testing system |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2300687A1 true CA2300687A1 (en) | 2001-08-23 |
Family
ID=4165513
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA 2300687 Abandoned CA2300687A1 (en) | 2000-02-23 | 2000-02-23 | Novel elisa testing system |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA2300687A1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021005629A1 (en) * | 2019-07-05 | 2021-01-14 | Da Broi Ugo | Portable spr sensor for detecting anaphylaxis, and corresponding method |
-
2000
- 2000-02-23 CA CA 2300687 patent/CA2300687A1/en not_active Abandoned
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2021005629A1 (en) * | 2019-07-05 | 2021-01-14 | Da Broi Ugo | Portable spr sensor for detecting anaphylaxis, and corresponding method |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request | ||
| FZDE | Dead |