Direct chemiluminiscence kit for detecting beta-hCG content and use method of direct chemiluminiscence kit

A chemiluminescence reagent, a direct technology, applied in the field of kits, can solve the problems of unstable system and increased variation of test values, and achieve the effect of saving research and development costs, reducing costs, and strong applicability

Pending Publication Date: 2021-03-30
BEAVERNANO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For a higher signal-to-noise ratio, some surfactants are generally added to the excitation solution to enhance the signal, but t...

Method used

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  • Direct chemiluminiscence kit for detecting beta-hCG content and use method of direct chemiluminiscence kit
  • Direct chemiluminiscence kit for detecting beta-hCG content and use method of direct chemiluminiscence kit
  • Direct chemiluminiscence kit for detecting beta-hCG content and use method of direct chemiluminiscence kit

Examples

Experimental program
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Effect test

preparation example Construction

[0043]The biotinylated β-hCG antibody label is prepared by the following steps: add 500 μg of β-hCG antibody to an ultrafiltration tube at 4°C, centrifuge at 12,000 rpm, 10 min / time, repeat three times, remove the original buffer, add Put 500μL LPBS in an ice bath; dissolve 3.1mg sulfo-NHS-Biotin in 700μL anhydrous DMF to obtain a 10mM Biotin / DMF solution; add 16.7μL 10mM Biotin / DMF solution to the antibody solution, mix quickly, and place on ice React for 2 hours; after the reaction, centrifuge the reaction solution at 4°C at 12,000 rpm for 10 min / time, repeat three times to remove unreacted and hydrolyzed biotin and other small molecules; add 500 μL PBS, pipette repeatedly for 10 min, take it out, and use 50 μL per tube Aliquot and store at 4°C.

[0044] The acridinium ester-labeled β-hCG antibody method specifically includes the following steps:

[0045] The method for labeling acridinium ester with β-hCG antibody is as follows: add 500 μg antibody to the ultrafilt...

example 1

[0053] Example 1: β-hCG antibody labeled with biotin

[0054] Reagent consumables:

[0055] 1.PBS: 10mM PBS, pH7.4

[0056] 2. Anhydrous DMF (remove water one day before use)

[0057] 3. Sulfo-NHS-Biotin (BBI Life sciences) C100213 50mg powder

[0058] 4. Monoclonal Antibody to β-hCG: Meridian product number E86308M concentration 5.3mg / ml

[0059] 5. Ultrafiltration tube Millpore Ultra-4MWCO is 50KD. Ultrafiltration tubes need to be pre-washed with sterile PBS before use. Once wet, the filter membrane in the ultrafiltration centrifuge tube should be prevented from drying out. If not used immediately after prewashing, allow liquid to remain on filter until use.

[0060] 6. Molecular sieve, type 5A, 3-5mm.

[0061] Experimental steps:

[0062] 1. Pre-wash the ultrafiltration tube several times with sterile PBS. Turn on the ice maker ahead of time and prepare the ice box. Take out Sulfo-NHS-Biotin from -20°C in advance and equilibrate to room temperature to prevent ...

example 2

[0068] Example 2: β-hCG antibody labeling acridinium ester

[0069] Reagent Consumables

[0070] 1. Coupling buffer: 0.05mol / L CBS (Na 2 CO 3 -NaHCO 3 buffer).

[0071] 2. Anhydrous DMF (it is best to remove water one day before use), and the method of removing water refers to the method of using molecular sieves.

[0072] 3. Acridine ester (NSP-DMAE-NHS): Suzhou Yake product number Y0266, 10mg / tube.

[0073] 4. Monoclonal Antibody to β-hCG: Meridian product number E86308M concentration 5.9mg / ml

[0074] 5. Ultrafiltration tube Millpore Ultra-4MWCO is 50KD. Ultrafiltration tubes need to be pre-washed with sterile PBS before use.

[0075] Once wet, the filter membrane in the ultrafiltration centrifuge tube should be prevented from drying out. If not used immediately after prewashing, allow liquid to remain on filter until use.

[0076] 7. Molecular sieve, type 5A, 3-5mm.

[0077] Experimental procedure

[0078] 1. Prepare sterile CBS, pre-wash the ultrafiltr...

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Abstract

The invention discloses a magnetic particle direct chemiluminiscence kit for detecting the content of free beta-hCG (human chorionic gonadotropin) and a use method of the magnetic particle direct chemiluminiscence kit. The kit comprises magnetic microspheres coated with streptavidin, a biotin-labeled beta-hCG antibody, a beta-hCG standard substance, an acridinium ester-labeled antibody with different beta hCG recognition sites, a washing buffer solution, an acridinium ester labeled antibody diluent, a standard substance diluent, an excitation solution A and an excitation solution B containing1%-2% of Triton X100 and ethanol. According to the kit, the magnetic microsphere and streptavidin protein coupling technology, the biotin antibody labeling technology and the acridinium ester antibodylabeling technology are combined, and particularly components and proportions of the excitation solutions are debugged and improved, and therefore, the kit has various characteristics of high sensitivity, strong specificity, high accuracy and high repeatability. The kit can be used for detecting the direct chemiluminiscence performance of magnetic particles and better detecting the content of free beta-hCG (human chorionic gonadotropin).

Description

technical field [0001] The invention relates to the technical field of kits, in particular to a direct chemiluminescent kit for detecting β-hCG content and a use method thereof. Background technique [0002] Human chorionic gonadotropin (hCG) is a glycoprotein hormone secreted by trophoblast cells of the placenta, which is composed of two subunits α and β in a non-covalent bond. Glycoprotein hormone with a molecular weight of 36700. hCG is produced by the placenta and cytotrophoblast cells, and its α subunit has the same amino acid series as the α subunit of other hormones in the glycoprotein hormone family (such as LH, FSH, and TSH), while the β subunit is specific. [0003] In clinical testing, hCG is mainly used to diagnose early pregnancy, hydatidiform mole and ectopic pregnancy. The detection of β-hCG is mainly used for the screening of Down syndrome and as a tumor marker for tumor screening. The detection method of hCG is similar to that of β-hCG. Chemiluminescent i...

Claims

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Application Information

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IPC IPC(8): G01N33/76G01N33/543G01N21/76
CPCG01N33/76G01N33/54326G01N21/76G01N2333/59
Inventor 任辉万东菊
Owner BEAVERNANO TECH
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