Anti-Kratom monoclonal antibody and kit for detecting Kratom

A technology for cloning antibodies and anti-kratom monoclonals, which can be used in biological testing, measuring devices, and material inspection products. It can solve problems such as doping or contamination, and achieve high sensitivity, high affinity, and high specificity.

Active Publication Date: 2021-06-22
HANGZHOU CLONGENE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Adulteration or contamination with any co-swallowed substances is also a risk

Method used

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  • Anti-Kratom monoclonal antibody and kit for detecting Kratom
  • Anti-Kratom monoclonal antibody and kit for detecting Kratom
  • Anti-Kratom monoclonal antibody and kit for detecting Kratom

Examples

Experimental program
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Effect test

preparation example Construction

[0066] Preparation of card pain antigens, including the following steps:

[0067] Preparation of a fully antigen of a capumn wood base:

[0068] a) Protect the amino group: dissolve the cap column wood base in an organic solvent, stirred at room temperature, under an ice water bath, inert gas protection, slowly drop the caporial carnoid gas amount of 1-3 times trifluoroacetic anhydride, withdraw the ice Water bath, room temperature, stirring reaction 2-4h; After the reaction, the solvent was evaporated under reduced pressure in a water bath at 40-50 ° C to give a light yellow oil liquid, and 5-15 ml of n-hexane dissolved, crystallized into a white solid in an ice water bath. Hisherache is washed with white solid;

[0069]

[0070] b) Hydrolyzed ester group: Add 5-15 mL of methanol and 1-3 ml of water to the obtained white solid, then add 5-10-fold mole of lithium hydroxide, and continue to react 20h after stirring; the reaction is completed, with a thin plate Analysis, raw mater...

Embodiment 1

[0111] Example 1: Preparation of Cardiac Antigen

[0112] Preparation of a capillarkine (Antigen1) complete antigen:

[0113] a) Protect amino group

[0114] 398.5 mg (1 mmol) cap column wooden base powder was added to 150 ml of three flasks, 60 ml of dichloromethane, stirred at room temperature, under ice water bath, under nitrogen protection, slowly add 420 mg (2 mmol) trifluoroacetic anhydride, The ice water bath was removed, stirred at room temperature for 3 hours. After the reaction, the evaporation of the solvent was evaporated under a water bath at 45 ° C to give a pale yellow oily liquid, and 10 ml of n-hexane dissolved. Crystallization in an ice water bath can be crystallized into a white solid. 420 mg (0.86 mmol) of the white solid was washed with n-hexane.

[0115] b) hydrolyzate

[0116] 10 mL of methanol and 2 ml of water were added to the white solid, and 158 mg (6.88 mmol) lithium hydroxide was added, and the dissolution was continued for more than 20 hours.

[0117...

Embodiment 2

[0136] Example 2: Preparation of Cardial Antigen

[0137] Preparation of a capillarkine (Antigen1) complete antigen:

[0138] a) Protect amino group

[0139] 398.5 mg (1 mmol) capillaine powder was added to 150 ml of three flasks, 60 ml of dichloromethane was added, and the mixture was stirred at room temperature, under nitrogen protection, slowly add 210 mg (1 mmol) trifluoroacetic anhydride, The ice water bath was removed, stirred at room temperature for 3 hours. After the end, the solvent was evaporated under a water bath at 405 ° C to give a pale yellow oily liquid, and 10 ml of n-hexane dissolved. Crystallization in an ice water bath can be crystallized into a white solid. 239 mg (0.6 mmol) of the white solid was washed with n-hexane.

[0140] b) hydrolyzate

[0141] 10 ml of methanol and 2 mL of water were added to the white solid, and then 110 mg (4.8 mmol) lithium hydroxide was added, and the reaction was continued for more than 20 hours after stirring dissolution.

[0142...

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Abstract

The invention relates to the field of biological detection, and discloses an anti-Kratom monoclonal antibody and kit for detecting Kratom. The anti-Kratom monoclonal antibody disclosed by the invention is secreted by a hybridoma cell strain Kratom11B7, can be used for simultaneously detecting capillamine and 7-hydroxyl capillamine, and can be used for better detecting edible Kratom leaves, a Kratom leaf extract and derivatives thereof. The invention has the characteristics of high affinity, high specificity and high sensitivity. Compared with other products in the prior art, the kit provided by the invention has more advantages in the aspects of sensitivity, specificity, detection limit and the like when being used for detecting Kratom in saliva.

Description

Technical field [0001] The present invention relates to the field of biological detection, and more particularly to an anti-kinelette monoclonal antibody and a kit for detecting a cardine. Background technique [0002] Card pain, 科, is a spiritual active plant that grows in Southeast Asia, especially in Thailand and Malaysia. Fresh or dry card pain leaves can be consumed by suction, chew or tea. The low-dose card pain has a stimuli. For a long time, it has been used by Southeast Asia's physical workers as a stimulant to cancel the work fatigue; the high-dose card pain has a calm-anesthesia effect, so the card pain leaves are used as a traditional drug and opium alternative. [0003] The pharmacological function of cardiac pain is mainly achieved by alkaloid 7-hydroxyap column wooden base (formula II) and cap columns (formula I). Although the structure of these alkaloid molecules is similar, these substances do not have a distortion activity. Conversely, these alkals are mainly in...

Claims

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Application Information

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IPC IPC(8): C07K16/16G01N33/94G01N33/577G01N33/558G01N33/58
CPCC07K16/16G01N33/948G01N33/577G01N33/558G01N33/587C07K2317/92Y02P20/55
Inventor 郑智彪郑曙剑
Owner HANGZHOU CLONGENE BIOTECH
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