Homogeneous agglutination immunoassay method and kit for such method

An immunoassay and reagent technology, applied in the field of homogeneous agglutination assay, can solve the problems of easy access

Inactive Publication Date: 2012-03-28
BECKMAN COULTER BIOMEDICAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] A drawback of the known methods is that they require distinct epitopes on the antigen that are spatially accessible
This does not apply to all clinical analytes
Additionally, the need to use different antibodies in known assays represents a disadvantage

Method used

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  • Homogeneous agglutination immunoassay method and kit for such method
  • Homogeneous agglutination immunoassay method and kit for such method
  • Homogeneous agglutination immunoassay method and kit for such method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] 1) Reagent

[0073] The reagents used have the following composition:

[0074] i) Reagent A

[0075]

[0076] Join H 2 O up to 1000mL. Optionally, preservatives, stabilizers, EDTA or protein can be added to Reagent A. However, they are not required for the method according to the invention.

[0077] ii) Reagent B

[0078]

[0079] Join H 2 O up to 1000mL. Optionally, preservatives, stabilizers, EDTA, proteins, or detergents may be included in Reagent B. However, they are not required for the method according to the invention.

[0080] iii) Calibrator

[0081] A set of calibrators with a concentration range of 0-530mg / L CRP.

[0082] 2) Determination

[0083] The determination is carried out at 37° C. in an automatic analyzer Olympus AU2700. All the different urea concentrations of reagent A were used in combination with each particle size of reagent B.

[0084] - 2 μL of Calibrator was pipetted into 150 μL of Reagent A and mixed;

[0085] - After 3.2 m...

Embodiment 2

[0091] 1) Reagent

[0092] Select reagent A with a urea concentration of 8.0 mol / L. As in the case of Example 1, Reagent B can contain either 158 nm diameter particles or 101 nm diameter particles.

[0093] 2) Determination

[0094] Similar to Example 1, the entire assay was performed in an Olympus AU2700 analyzer at 37°C.

[0095] - 2 µL of each calibrator was pipetted into 150 µL of Reagent A and mixed;

[0096] - After 3.2 min, add 75 μL of Reagent B and mix.

[0097] - Then measure the OD value at 520nm, 600nm or 700nm during 4.9 minutes.

[0098] - For each wavelength, the change in OD value is expressed as the change between the beginning and end of the measurement period.

[0099] -For each wavelength, the change in OD is plotted against the concentration of CRP in a single calibrator ( Figure 4 (158nm); Figure 5 (101nm)).

[0100] figure 2 and image 3 The measurement results of Example 1 using 2 different particle sizes at different urea concentrations ar...

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Abstract

A homogeneous agglutination immunoassay method wherein a sample possibly containing the analyte to be measured is mixed with a binding partner for the analyte and with at least one chaotropic agent, monovalent ions in form of at least one salt, or with a combination of at least one chaotropic agent and monovalent ions in form of at least one salt, measuring at least one signal related to interactions of the binding partner with the analyte over a reaction time and calculating from at least one of the signals measured the concentration of analyte in the sample.

Description

technical field [0001] The present invention relates to a method for a homogeneous agglutination assay and a kit for this method. Background technique [0002] Homogeneous agglutination assays are commonly used to determine the concentration of an analyte in a sample by exploiting the binding reaction between specific immunopartners for analyte detection and quantification. In such assays, the degree of agglutination due to aggregate formation is measured and used as the basis for analyte calculations. As a principle in homogeneous assays, the sample to be analyzed is mixed with the binding partner and, after an optional incubation period, the agglutination in the assay is measured directly without any necessary separation steps. [0003] Typically, the binding partner is an antibody specific for the analyte of interest. Suitable analytes must expose more than two homogeneous or heterogeneous epitopes for binding partners. Of course, antigens can also be used as binding p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543
CPCG01N33/54346G01N33/54313
Inventor 马丁・伯克哈特
Owner BECKMAN COULTER BIOMEDICAL
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