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Cocaine-and amphetamine-regulated transcript peptides single-chain antibody and application thereof

A technology for regulating transcription and single-chain antibodies, applied in the direction of antibodies, peptides, specific peptides, etc.

Inactive Publication Date: 2011-11-09
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There is no literature report on the screening of antibodies for the treatment of addictive behaviors caused by psychoactive substances such as cocaine (including opioids, cocaine, amphetamine, and ketamine) by phage display technology

Method used

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  • Cocaine-and amphetamine-regulated transcript peptides single-chain antibody and application thereof
  • Cocaine-and amphetamine-regulated transcript peptides single-chain antibody and application thereof
  • Cocaine-and amphetamine-regulated transcript peptides single-chain antibody and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] pGEX-4T3-CART 55-102 (Lin Qin, Cocaine-amphetamine-regulated transcription peptide antibody inhibits the formation and reconstruction of morphine-induced mouse conditioned place preference, 2006, Master's thesis of Second Military Medical University, pGEX-4T3 was purchased from Amersham phaemacia company) expressing glycerol bacteria 30μl transfer In 3ml of LB medium containing 100μg / ml, cultivate overnight at 37°C on a shaker; add 1ml of the transfer bacteria solution to 100ml of LB medium containing 100μg / ml Amp, grow on a shaker at 37°C until the OD600 value is 0.4, add 1M IPTG 400μl to make the final concentration 0.4mM, 37°C, 230r.p.m. shaker to induce expression for 5h, 4°C low-temperature centrifuge at 12,000r.p.m. for 20min to collect the bacteria, add PBS to suspend the bacteria at 10ml / wet weight (g) . Suspend the bacteria overnight in an ice bath, sonicate at 100W for 1 second, rest for 1 second, and sonicate for 20 minutes. Centrifuge at 12,000r.p.m. at 4℃...

Embodiment 2

[0035] Example 2: Effect of CART single-chain antibody (AH36) on cocaine-induced drug dependence

[0036] The antibody was used in animal experiments, and it was confirmed that the cocaine-amphetamine regulatory transcription peptide antibody reduces the drug dependence of the body caused by the use of cocaine and other psychotropic drugs. The result is as follows:

[0037] (1) Behavioral Sensitization Model

[0038] 1. Spontaneous Activity Recording Devices

[0039] The conditional place preference shuttle box black box used in this experiment was used as the spontaneous activity recording device. The black box is placed in a 100×100×100 soundproof box with an inner diameter of 15×15×35cm, and a piece of smooth barbed wire is placed 5cm above the bottom of the box; the shuttle door is closed. The observation box has the effect of light insulation and sound insulation, and the sound intensity attenuation is 20-25dB. On the top of the box, white lighting and infrared LEDs a...

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Abstract

Disclosed is a new antibody which can be used to relieve addiction caused by nerve active substances, including opium, cocaine, benzedrine, ketamine, etc. Cocain and amphetamine-regulated transcript peptide antibody gene is selected through phage display technology; then single-chain antibody sequence of the cocain and amphetamine-regulated transcript peptide(CART) and the procaryon expression vector are connected and are induced to express and to be purified to obtain single-chain antibody in response to CART, which is named as CARTscFv(AH1,AH4B,AH6,AH19,AH33B,AH36). Injecteion of CARTscFvinto the abdominal cavity of a mouse, together with the CART at the peripheral circulation system, leads to the reduction of the CART in the central nervous system of the mouse. Animal model with behavioral sensitization induced by cocain proves that the CART single-chain antibody can restrain the expression of the behavioral sensitization of the mouse induced by cacain, which shows that the CART single-chain antibody is of great potential in curing addiction and particularly relapse caused by nerve active.

Description

Technical field: [0001] The invention relates to the technical field of biomedical engineering, in particular to the preparation and application of an antibody capable of resisting mental dependence (relapse) caused by psychoactive substances such as cocaine. Background technique: [0002] Psychoactive substance dependence (drug abuse) and relapse are serious problems in today's society. At present, there is no ideal method to prevent and treat relapse after detoxification. In recent years, the research on cocaine-amphetamine-regulated transcription peptide (CART) has been deepened in the scientific community. Cocaine-amphetamine-regulated transcription peptide (CART) is a class of peptides related to drug addiction and dependence effects. The substance can cause significant expression of cocaine-amphetamine-regulated transcription peptide, but its receptor has not been successfully cloned so far. Increasing the level of CART in the ventral dorsal tegmental area (VTA), a br...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/44A61K39/395A61P25/36A61P25/30
Inventor 由振东路长林柴景蕊
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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