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A kind of latex-based melamine rapid detection method and kit

A melamine and latex technology, applied in the field of interdisciplinary, can solve the problems such as the inability to obtain products, and achieve the effects of low reagent production cost, simple preparation and short time consumption

Active Publication Date: 2016-04-20
BEIJING PURKINJE GENERAL INSTR
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the ligand to be coupled also contains carboxyl groups (e.g. proteins, such as antibodies), it may be activated by EDC, leading to aggregation between the ligands, so that the desired product cannot be obtained

Method used

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  • A kind of latex-based melamine rapid detection method and kit
  • A kind of latex-based melamine rapid detection method and kit
  • A kind of latex-based melamine rapid detection method and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Embodiment 1: the preparation of buffer R1

[0063] 40mM, PH7.4 PBS preparation: ①Weigh 0.0624g of sodium dihydrogen phosphate, and dilute to 100ml with ultra-pure water; ②Weigh 7.16g of disodium hydrogen phosphate, and dilute to 500ml with ultra-pure water; Sodium 0.14g, add 3.8ml of reagent ① and 16.2ml of reagent ②.

[0064] This is 40mM phosphate buffered saline (PBS), pH 7.4.

[0065] Preparation of R1:

[0066] PEG60000.3g

[0067] Sodium azide 0.01g

[0068] Tween-2020ul

[0069] After weighing various chemicals, dilute to 10ml with 40mM, pH7.4 PBS.

[0070] Since PEG6000 does not dissolve at room temperature, place the reagent in a 60-degree water bath to heat it, and keep stirring to mix it evenly. The dissolved reagent is a colorless transparent liquid.

[0071] After preparing the reagents, store them in a refrigerator at 2-8°C, and the shelf life is 12 months.

Embodiment 2

[0072] Example 2: Reagents for preparing and preserving latex microspheres coated with melamine antibody

[0073] Activation buffer:

[0074] Preparation of 10 mM PBS (pH 7.4): the 40 mM PBS prepared in Example 1 was diluted four times, and then 0.70 g / L Tween-20 was added to the obtained 10 mM PBS.

[0075] Reconstitution solution:

[0076] Glycine 11.3g / L

[0077] Sodium azide 1.0g / L

[0078] Sorbitol 10.0g / L

[0079] BSA5.0g / L

[0080] EDTA·Na22.0g / L

[0081] First weigh various solid compounds, and then use ultra-pure water to make up the volume. In order to increase the dissolution rate, it can be placed in a 37-degree incubator. Finally, add sodium hydroxide solution (20%) to adjust the pH to 8.00±0.08. The prepared reagent is a colorless, clear and transparent liquid. The prepared complex solution can be stored in a refrigerator at 2-8°C, and the shelf life is 12 months.

[0082] Quenching solution:

[0083] Glycine 75g / L

[0084] After weighing glycine, dilu...

Embodiment 3

[0088] Embodiment 3: the preparation technology of the latex microsphere coated with melamine antibody

[0089] The reaction system is 100ul.

[0090] activation

[0091] Take a 1.5ml centrifuge tube, add 70ul of activation buffer, and use a pipette to take 10ul of polystyrene latex microspheres (purchased from PolyMicrospheres, USA, particle size 130-170nm, carboxyl content 160-170μeq / g) and centrifuge here In the tube, keep pipetting to disperse the latex particles evenly into the activation buffer, so as to obtain a white emulsion.

[0092] Weigh solid carbodiimide hydrochloride (EDCA; purchased from Shanghai Covalent Chemical Technology Co., Ltd.) and dissolve it in the activation buffer to a concentration of 0.001 mg / ul. Add 10 μl of EDCA solution to the above-mentioned latex particle buffer, and activate at room temperature for 5 minutes. The activated latex system is still a white emulsion, but the viscosity has decreased.

[0093] coupling

[0094] Add the activat...

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Abstract

The invention discloses a melamine antibody-coated latex microsphere solution, a preparation method of the melamine antibody-coated latex microsphere solution, a kit containing the melamine antibody-coated latex microsphere solution, and a method for detecting melamine in a sample by the melamine antibody-coated latex microsphere solution.

Description

technical field [0001] The invention relates to the interdisciplinary fields of immunology, medicine, biology and food safety, and more specifically relates to a method for detecting melamine based on polystyrene latex microspheres. Background technique [0002] In the food industry, the nitrogen atom content is determined by the Kjeldahl method (GB / T5009.5-2010; the first method) to indirectly calculate the protein content. In recent years, some unscrupulous traders have added chemical raw material melamine to food or feed in order to increase the apparent protein content determined by the Kjeldahl method. Melamine can cause stones in the human urinary system, such as bladder stones and kidney stones, and is therefore strictly prohibited from being added to food. [0003] At present, the detection methods of melamine include high-performance liquid chromatography, liquid chromatography-mass spectrometry, gas chromatography-mass spectrometry, etc. Although these methods hav...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577
CPCG01N21/31G01N33/53G01N33/531
Inventor 王琳张小波郑清林
Owner BEIJING PURKINJE GENERAL INSTR