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Application of anti-camk2a autoantibody reagents in preparation of kits for diagnosing neurological symptoms related diseases

A technology of autoantibodies and kits, applied in the fields of resistance to vector-borne diseases, disease diagnosis, biochemical equipment and methods, etc.

Active Publication Date: 2022-08-09
SHAANXI MYBIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, although progress has been made in identifying autoantibody-mediated immune mechanisms in patients with autoimmune encephalitis, there are still a proportion of patients who exhibit encephalitic symptoms with negative serological test results. Therefore, identification of autoantibody-binding Neoantigens, improving existing diagnostics and therapeutics, remain a daunting task

Method used

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  • Application of anti-camk2a autoantibody reagents in preparation of kits for diagnosing neurological symptoms related diseases
  • Application of anti-camk2a autoantibody reagents in preparation of kits for diagnosing neurological symptoms related diseases
  • Application of anti-camk2a autoantibody reagents in preparation of kits for diagnosing neurological symptoms related diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] The immune fluorescent method of rats primary neuron cells

[0066] Step 1 Rat Primary Neurar cell separation

[0067] 1. 10 % hydrolytic chloride anesthesia mater, 75 % alcohol is soaked and disinfected for 3min, the biological safety cabinet is taken out of the tire mouse;

[0068] 2. Use the pre -cold HBSS (137mm naCl, 5.4mm kCl, 0.6mm MGSO 4 · 7H 2 O, 0.5mm mgCl 2 · 6H 2 O, 0.3mm na 2 HPO 4 · 2H 2 O, 0.4mm kh 2 PO 4 , 5.6mm glucose, 4.2mm nahco 3 , PH7.2 ~ 7.4) Rinse the tire rat twice, dissecting the tire brain in the pre -cold HBSS in the pre -cold DMEM;

[0069] 3. Discard DMEM, cut the tissue block into 1M with ophthalmology 3 Size mixture. Add papaya protease (concentration is 2 mg / ml). 30 minutes of digestion at 37 ° C in 10cm dish, gently shake every 5min. Generally, 10 to 15 brain tissues are 20ml for gastrointestinal liquid;

[0070] 4. After the digestion is over, move the cells to a 50ml centrifugal tube, add 20ml DMEM to re -suspension, 1 mL gun head 10 time...

Embodiment 2

[0096] Camk2a's immune sedimentation

[0097] There are three serum used in the immune sedimentation experiments of this embodiment, namely the serum of patients 1, health subject 1, and serum of healthy subjects 2, which obtain a total of three to eluted liquids. The specific experimental steps are as follows:

[0098] 1. Take the primary neuron cells of the rats of 6 rats, abandon the clearing, wash 2 times PBS, 0.4 % multi -polyethylene formaldehyde fixed for 10 minutes, abandon the clearing, wash 1 × hepes 3 times;

[0099] 2. Serum incubation: Take 10UL serum to 10ml DMEM (diluted 1: 100), filter the filtering fluid to the fixed cells with 0.22um filter membrane, and incubate for 2h by room temperature;

[0100] 3. Take 15ul Ge's Proteina and add it to a 2ml centrifugal tube. 2 HPO 4 Wash 150mmnacl) 3 times, close 2h with 300ul 4 % BSA;

[0101] 4. Place the incubated cells on the ice, abandon the clearing, wash the PBS 2 times, add 500UL cracking liquid (150mm NaCl, 1mm EDTA...

Embodiment 3

[0109] Immunization

[0110] 1. Electricity: Take the samples of Step 7 for Practice Example 7 for SDS-PAGE;

[0111] 2. Transfer: After the electrophoresis is completed, use the wet method to turn the film.

[0112] 3. Closed: Use 5 % skim milk powder room temperature to close 1h;

[0113] 4. One anti -incubation: incubation anti -Camk2a's polyxlon antibody (antibody purchased from Sigma) at a ratio of 1: 2000, and incubation for 2h;

[0114] 5. Washing: TBST 3 times, 5min each time; each time;

[0115] 6. Second resistance incubation: add the two resistance of HRP marked, room temperature incubation 1h

[0116] 7. Wash TBST 3 times, 5min each time; each time;

[0117] 8, color rendering: add chemical luminous fluid to record results.

[0118] Experimental results:

[0119] image 3 The WB identification results of serum immunization sediment, where M: Marker; 1: The sample above is the patient's serum immunization sediment; It can be seen from Figure 3 that there are bands that ...

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Abstract

The invention relates to the application of an anti-CAMK2A autoantibody reagent in the preparation of a kit for diagnosing diseases related to symptoms of the nervous system, and belongs to the technical field of disease diagnosis kits. The invention provides the application of an anti-CAMK2A autoantibody reagent in preparing a kit for diagnosing diseases related to symptoms of nervous system. The kit prepared by the application of the present invention can realize the detection of CAMK2A autoantibody, and can be used to diagnose nervous system diseases, especially nervous system diseases related to one or more symptoms, and the symptoms include sleep disturbance, lethargy , depression, cognitive and psychiatric disorders, autonomic dysfunction, seizures, decreased level of consciousness, confusion. The kit prepared by the application of the present invention can also be used to distinguish autoimmune diseases, especially autoimmune diseases and non-autoimmune diseases of the nervous system.

Description

Technical field [0001] The invention involves the technical field of the disease diagnostic kit, which specifically involves the application of the reagent of the anti -Camk2a antibody in the institution box that prepares the diagnostic nervous system symptoms. Background technique [0002] Autoimmune Encephalitis (AE) refers to a type of encephalitis mediated by autoimmune mechanism. It is a central nervous system inflammation of the central nervous system mediated by antibodies and causing neuronal structural dysfunction. Its antibodies include Antigen antibody antigen (producing sub -tumor syndrome) and antigening antigen antibodies of antigening antigen (producing sub -tumor syndrome). [0003] Existing literature reports and clinical testing of autoimmune encephalitis antibody spectrum includes: N-methyl-D skydamine receptor (NMDAR), α-amino-3 hydroxyl-5thyl-4 isozolizole Receptor (AMPAR), neuroplasticoma-rich neuroplastinoma (LGI1), contacted protein-related protein-2 (CASP...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/573C12N9/12
CPCG01N33/573C12N9/1205G01N2800/28G01N2800/24G01N2333/9121Y02A50/30
Inventor 闫亚平任妮柴单单曹玉妍封雪李科
Owner SHAANXI MYBIOTECH CO LTD
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