Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Agglutination assay

An analysis method and analyte technology, applied in the field of agglutination analysis, can solve problems such as difficult electronic recording of data, loss of sensitivity, damage to efficacy, etc., and achieve the effects of improving needs, enhancing sensitivity, and simplifying manufacturing

Inactive Publication Date: 2008-11-12
PLATFORM DIAGNOSTICS LTD
View PDF21 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, visual endpoints are subjective and difficult to record data electronically
[0017] An important consideration is that, by and large, these agglutination-based assays are limited to the detection of large analytes with multiple epitopes that form large and stable agglutinates
For smaller analytes with few epitopes, or with analytes with a limited number of available epitopes, their potency can be compromised due to the reduced number of binding events, possibly resulting in weakened aggregates and loss of sensitivity

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Agglutination assay
  • Agglutination assay
  • Agglutination assay

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0130] Embodiment 1. Preparation of soluble central reagent 1

[0131] 1. Using a 1.6×15 cm G25M Sephadex column, desalt anti-hCG (α subunit) into 0.1 M phosphate buffer at pH 7.5, and measure the concentration and yield.

[0132] 2. Anti-hCG antibody was activated using 8 molar equivalents of NHS-PEG-MAL. The reaction mixture was incubated at 20°C for 2 hours. The reaction was quenched with 100 molar equivalents of glycine, and maleimide-activated anti-hCG was desalted into 5 mM EDTA, PBS buffer, pH 7.3, using two passes through a 1.6 x 15 cm G50F Sephadex column. Determine the concentration and yield of activated antibody.

[0133] 3. Activation of 500KD aminodextran with 1000 molar equivalents of 2-iminothiol (2-IT). The reaction mixture was incubated at 20°C for 110 minutes. Thiol-activated aminodextran was desalted into 5 mM EDTA, pH 7.3, PBS buffer using G25M Sephadex medium. Thiol:aminodextran incorporation was determined by Ellman analysis.

[0134] 4. Add 25 m...

Embodiment 2

[0135] Embodiment 2. Preparation of soluble central reagent 2

[0136] 1. Using a 1.6×15 cm G25M Sephadex column, desalt anti-hCG (α subunit) into 0.1 M phosphate buffer at pH 7.5, and measure the concentration and yield. .

[0137] 2. Anti-hCG antibody was activated using 8 molar equivalents of NHS-PEG-MAL. The reaction mixture was incubated at 20°C for 2 hours. The reaction was quenched with 100 molar equivalents of glycine, and maleimide-activated anti-hCG was desalted into 5 mM EDTA, PBS buffer, pH 7.3, using two passes through a 1.6 x 15 cm G50F Sephadex column. Determine the concentration and yield of activated antibody.

[0138] 3. Activate a 500KD aminodextran with 1000 molar equivalents of 2-iminothiol (2-IT). The reaction mixture was incubated at 20°C for 110 minutes. Thiol-activated aminodextran was desalted into 5 mM EDTA, pH 7.3, PBS buffer using G25M Sephadex medium. Thiol:aminodextran incorporation was determined by Ellman analysis.

[0139] 4. Add 25 m...

Embodiment 3

[0141] Example 3. Preparation of membrane strips for testing using wet reagents

[0142] Membrane material is cut to the following dimensions:

[0143] (i) Liquid absorbent core, such as Surewick G028-14 (Millipore), 30mm×60mm.

[0144] (ii) Condensate trapping membrane, eg Fusion 5 (Whatman), 5mm x 60mm.

[0145] (iii) Absorbent trough, eg Absorbent Pad 222 (Ahlstrom), 55mm x 60mm.

[0146] (iv) Self-adhesive plastic (×2)

[0147] For example 0.04" Clear polyester (G&L) with D / C hydrophilic PSA 70mm x 100mm.

[0148] Composite "cards" of the above materials are assembled as shown in Figure 1. To ensure good liquid transport between successive sections of the strip, adjacent membrane material overlapped by approximately 1 mm. A second sheet of self-adhesive plastic is firmly applied to the upper surface. The "cards" showing the results were sliced ​​into 5 mm strips and the plastic trimmed to allow reagent and sample access to the wick.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to agglutination assays and related kits, reagents and devices. In particular methods of assaying small analytes having few epitopes are disclosed, by means of using hub moieties to which multiple analytes may be bound by a first epitope, together with a further moiety capable of binding a second analyte epitope and which is also capable of binding to a detectable particle. Stable agglutinated complexes may be so formed, which may used as the basis for various assay formats.

Description

field of invention [0001] The present invention relates to an agglutination assay for detecting an analyte in a sample. In particular, the present invention relates to assays and assay devices based on porous supports, kits comprising means for performing such assays, and assay methods for detecting analytes in samples. Background of the invention [0002] Immunoassays are well-established techniques for the detection and quantification of analytes in samples. They are particularly suitable for detecting and / or measuring substances in biological samples as an aid in the diagnosis and prognosis of diseases, and for predicting a patient's response to treatment. Technologies that have revolutionized diagnostic medicine, such as radioimmunoassays and enzyme immunoassays, are based on the detection of antibody-antigen interactions. A number of detection systems are available, including the use of radioactive or enzyme-labeled antigens, antibodies or complexes thereof. Many req...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N33/58G01N33/558G01N33/543G01N33/53G01N33/548
CPCG01N33/54313G01N33/54306G01N33/54393G01N33/544G01N33/552G01N33/555
Inventor 卡罗林·珍妮弗·拉德尔杰拉德·约翰·阿兰道格拉斯·罗伯特·艾文斯伊丽莎白·加纳尔
Owner PLATFORM DIAGNOSTICS LTD
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com