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Brain-derived neurotrophic factor determination kit, application thereof and brain-derived neurotrophic factor specific antibody

A neurotrophic factor and kit technology, used in growth factors/inducing factors, biological testing, neuregulin, etc., can solve the problems of difficult clinical trials and the lack of brain-derived neurotrophic factor assay kits. The effect of good diagnostic sensitivity

Pending Publication Date: 2022-06-24
深圳市安群生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Although there are currently preliminary studies on the use of BDNF in the auxiliary diagnosis and treatment judgment of depression, since there is no objective standard for the diagnosis of depression, clinical trials are difficult, and there is no brain-derived neurotrophic factor for the diagnosis of depression. Assay Kit

Method used

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  • Brain-derived neurotrophic factor determination kit, application thereof and brain-derived neurotrophic factor specific antibody
  • Brain-derived neurotrophic factor determination kit, application thereof and brain-derived neurotrophic factor specific antibody
  • Brain-derived neurotrophic factor determination kit, application thereof and brain-derived neurotrophic factor specific antibody

Examples

Experimental program
Comparison scheme
Effect test

preparation example 1

[0055] Preparation Example 1: Synthesis of BDNF-specific epitope peptides (1) and (2)

[0056] Tyr-Glu-Trp-Val-Thr-Ala-Ala-Asp-Lys-Lys-Thr-Ala-Val-Asp-Met-Ser (1)

[0057] Tyr-Lys-Lys-Arg-Ile-Gly-Trp-Arg-Phe-Ile-Arg-Ile-Asp-Thr-Ser (2)

[0058] 1. Raw materials used:

[0059] HMP resin (P-hydroxymethylphenoxymethyl polyethylene resin, purchased from sigma company)

[0060] Fmoc-AA (9-fluorenylmethoxycarbonyl protected amino acid, purchased from Merck)

[0061] NMP (nitrogen methyl pyrrolidone, purchased from sigma company)

[0062] DCM (dichloromethane, purchased from Zhongyuan Chemical Company)

[0063] MeoH (methanol, purchased from Zhongyuan Chemical Company)

[0064] Piperidine (piperidine, purchased from sigma)

[0065] DMAP (dimethylaminopyridine, purchased from sigma)

[0066] HOBT (Hydroxybenzotriazole, purchased from sigma)

[0067] DCC (dicyclohexylcarbodiimide, purchased from sigma)

[0068] TFA (trifluoroacetic acid, purchased from sigma)

[0069] EDT (1,...

preparation example 2

[0113] Preparation Example 2: Preparation of BDNF-specific Antibody

[0114] 1. Preparation of antigen:

[0115] The BDNF antigen was prepared by linking the BDNF-specific epitope peptide with the carrier protein KLH (keyhole limpet hemocyanin) by the method of Bis-diazotizedbenzidine dichloride (BDB).

[0116] Take 20.0 mg of BDNF-specific epitope peptide (1) or (2) prepared in Preparation Example 1, dissolve with 0.2 M borate buffer (pH 9.0); take 7.36 ml of KLH 6.25 mg / ml, cool to 0°C , take 1ml of BDB, put the two in an ice-water mixture, mix and avoid light, react on a shaker for 1-1.5h, adjust the pH to 9.0 with 0.2NaOH after the reaction is completed, dialyze overnight, and then divide into packages and store at -20°C.

[0117] The formula of borate buffer is: 80ml of 0.05mol / L borax, mixed with 20ml of 0.2mol / L boric acid.

[0118] 2. Immune animals to prepare monoclonal antibodies:

[0119] 2.1. After fully mixing the BDNF antigen (immunogen) prepared above with an...

Embodiment 1

[0128] Example 1: Preparation of chemiluminescence kit for BDNF

[0129] 1. Preparation of Magnetic Separation Reagent (Immune Magnetic Bead Working Solution Coated with BDNF Monoclonal Antibody 1)

[0130] (1) Initial washing and activation of magnetic beads

[0131] Take 200ul of magnetic beads, an aqueous dispersion of magnetic beads (purchased from Wuxi Biomag Biotechnology Co., Ltd.) with a content of 1 (w / v)%, and adsorb and precipitate with a magnetic base; discard the supernatant and add 0.8 ml of initial washing buffer solution (50mM 2-(N-morpholino)ethanesulfonic acid (MES aqueous solution, pH 6.5), ultrasonically (240W) and mixed for 4-5S, adsorb the precipitation, discard the supernatant, repeat the above process 3 times, Add the mixed solution of the carbodiimide solution of 750ul 10mg / ml and the N-hydroxysuccinimide solution of 10mg / ml, place on the thermostatic shaker for 15 minutes, thereby activate the carboxyl magnetic beads. Activation is completed, and ult...

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Abstract

The invention discloses a BDNF (brain-derived neurotrophic factor) determination kit. The BDNF determination kit comprises a solid-phase carrier, a BDNF specific antibody, a marker and a BDNF calibrator. The invention also discloses application of the kit provided by the invention in preparation of products for diagnosing depression. The invention further provides a BDNF specific antibody, the BDNF specific antibody is prepared by coupling the BDNF specific antigen epitope peptide with a carrier protein and then immunizing an animal, and the amino acid sequence of the BDNF specific antigen epitope peptide is one of the following two sequences: Tyr-Glu-Trp-Val-Thr-Ala-Ala-Asp-Lys-Lys-Thr-Ala-Val-Asp-Met-Ser (1) or Tyr-Lys-Lys-Arg-Ile-Gly-Trp-Arg-Phe-Ile-Arg-Ile-Asp-Thr-Ser (2). The kit prepared by the invention needs to have good diagnostic sensitivity, specificity and repeatability, and meets clinical application requirements.

Description

technical field [0001] The invention relates to the field of biology, in particular to a brain-derived neurotrophic factor (BDNF) assay kit and application of the kit in preparing a product for diagnosing depression and a brain-derived neurotrophic factor-specific antibody. Background technique [0002] Depression is an affective disorder that seriously affects physical and mental health. It is mainly characterized by persistent low mood, decreased interest, and slow thinking. Its lifetime prevalence is about 17%, and about 350 million people worldwide are affected by it. influences. According to the World Health Organization, by 2030 depression will rise to the first place in the total global disease burden. The latest data in 2019 show that the lifetime prevalence of depression in my country has reached 3.4%. Based on this, it is estimated that about 44 million people suffer from the disease. More importantly, the disease often recurs repeatedly, and the suicide rate is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/577G01N33/543G01N33/535G01N33/533C07K14/475C07K1/16C07K1/04C07K16/22
CPCG01N33/74G01N33/577G01N33/54326G01N33/54346G01N33/535G01N33/533C07K14/4756C07K16/22G01N2333/4756G01N2800/304
Inventor 朱建安朱仕杰
Owner 深圳市安群生物工程有限公司
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