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Method for increasing anti matrix effect in immunity detection for environmental sample, and dedicated buffer solution

A buffer solution, immunodetection technology, applied in the field of environmental sample immunodetection, to achieve the effect of enhancing stability, improving stability and consistency

Active Publication Date: 2007-12-26
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, in the immunoassay research of environmental samples, the interference law of complex matrix in environmental samples on immunoassay and the method of eliminating interference have not yet been developed.

Method used

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  • Method for increasing anti matrix effect in immunity detection for environmental sample, and dedicated buffer solution
  • Method for increasing anti matrix effect in immunity detection for environmental sample, and dedicated buffer solution
  • Method for increasing anti matrix effect in immunity detection for environmental sample, and dedicated buffer solution

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1. Obtaining of monoclonal antibody MC8C10 against microcystin-LR

[0067] 1. Obtaining the hybridoma cell line MC8C10 CGMCC No.2101

[0068] 1. Synthesis of Microcystin-LR Complete Antigen MC-LR-BSA

[0069] Microcystin-LR was chemically modified with 2-mercaptoethylamine, and an active group—amino group was introduced into its seventh amino acid (Mdha), and then the modified microcystin was synthesized by glutaraldehyde method. -LR (MC-LR) was coupled with bovine serum albumin, and the complete antigen was obtained after filtration chromatography, and the coupling ratio of the complete antigen was determined by MALDI-TOF / MS. The specific method is as follows:

[0070] (1) Amino modification of Microcystin-LR

[0071] ① Fully mix 2-mercaptoethylamine and MC-LR in an alkaline carbonate buffer (pH=8.0) according to a molar ratio of 3000:1; shake the mixture well and react at 50°C for 1.5 hours;

[0072] 2. After the reaction is completed, drop to room tempera...

Embodiment 2

[0118] Embodiment 2, using special buffer solution 1 to improve environmental sample immunoassay anti-matrix effect

[0119] The special buffer solution 1 used is PBS (configuration method: KCl 2.0g; KH 2 PO 4 2.4g; Na 2 HPO 4 12H 2 O 29g; add double distilled water to 1000mL. ), add the following substances a), b) and c): a) NaCl, to make the final concentration 50g / L; b) bovine serum albumin, to make the final concentration 10g / L; b) ethylene glycol Disodium amine tetraacetate, so that the final concentration is 5g / L.

[0120] 1. Eliminate the impact of pH in water on immunoassay

[0121] A competitive ELISA method was used to detect microcystin-LR in the water samples to be tested. Two treatments were set up in the experiment: antibody solution 1 and antibody solution 2 treatments.

[0122] The reagents and microtiter plates used were prepared according to the following methods:

[0123] 1. Preparation of water samples to be tested:

[0124] a. The HCl content in...

Embodiment 3

[0221] Embodiment 3, using special buffer solution 2 to improve the anti-matrix effect of environmental sample immunoassay

[0222] The special buffer solution 2 used is PBS (configuration method: KCl 1.0g; KH 2 PO 4 1.4g; Na 2 HPO 4 12H 2 O 13g; double-distilled water was added to 1000mL), and the following a), b) and c) were added: a) NaCl, to make the final concentration 10g / L; a) bovine serum albumin, to make the final concentration 2g / L; c) disodium ethylenediaminetetraacetic acid, so that the final concentration is 2g / L.

[0223] A competitive ELISA method was used to detect microcystin-LR in the water samples to be tested. Except that the special buffer solution was replaced by the special buffer solution 2, the experimental method and experimental materials were the same as steps 1, 2 and 3 of Example 2.

[0224] 1. Eliminate the impact of pH in water on immunoassay

[0225] The experimental result of adding common buffer solution is the same as step one of emb...

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Abstract

A special buffer solution of raising anti-substrate effect of environment sample immune-detection is prepared for adding NaCI as making its final concentration be 10-100g / L and adding inert protein as making its final concentration be 2-20g / L as well as adding complexon III as making its final concentration be 2-20g / L into existed buffer solution with concentration of 0.05-0.12mol / L and pH value of 7.0-8.0.

Description

technical field [0001] The invention relates to a method for improving the anti-matrix effect of environmental sample immunological detection and a special buffer solution thereof in the technical field of immunological detection of environmental samples. Background technique [0002] Matrix (Matrix) refers to the environment (Milieu) of an analyte (Analyte), that is, all components in the sample except the target analyte and their physical and chemical properties (such as viscosity, pH, temperature, etc.). [0003] Matrix Effect (Matrix Effect) is medically defined according to the clinical and laboratory standardization organization: (1) the influence of other components in the sample except the analyte on the measured value of the analyte; (2) the accurate determination of the matrix on the analytical method Interference in the ability of the analyte. [0004] Various matrices in environmental samples are quite different from those in other samples (such as blood samples...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N1/28
Inventor 何苗盛建武施汉昌
Owner TSINGHUA UNIV
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