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Detection material, preparation method and application of anti-gababr autoantibody in human body fluid

A 17T2A-GABABR, detection material technology, applied in the field of biomedicine, can solve the problems of large demand for antibodies, long detection time, cumbersome steps, etc., to achieve the effect of strong operation skills, short detection cycle, and high detection cost.

Active Publication Date: 2021-08-31
SHAANXI MYBIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of the problems existing in the prior art, the purpose of the present invention is to provide a detection material for detecting GABABR autoantibodies, a preparation method and application of the material, and solve the problems of time-consuming detection, cumbersome steps, and antibody demand of existing detection methods. Large size, high cost, low sensitivity, etc.

Method used

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  • Detection material, preparation method and application of anti-gababr autoantibody in human body fluid
  • Detection material, preparation method and application of anti-gababr autoantibody in human body fluid
  • Detection material, preparation method and application of anti-gababr autoantibody in human body fluid

Examples

Experimental program
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Embodiment 1

[0039]Step 1, plasmid construction: obtain the CDS sequence of GABABR as the target gene, insert the target gene with restriction sites into the 17T2A plasmid vector to obtain the recombinant plasmid vector 17T2A-GABABR, and extract the plasmid for subsequent experiments after the sequencing is correct. The plasmid vector construction of example specifically comprises the following steps:

[0040] Step 1.1: Obtain the CDS sequence of GABABR as the target gene by PCR method (artificial synthesis method is also optional), and add SalI / NotI restriction sites at both ends of the target gene;

[0041] Step 1.2: Insert the target gene with the restriction site into the 17T2A plasmid vector, the insertion site is SalI / NotI, to obtain a recombinant vector, which is named 17T2A-GABABR;

[0042] Among them, the 17T2A plasmid vector deletes the copGFP element on the basis of the pCDH-CMV-MCS-EF1-copGFP vector, and replaces the FE1 promoter with the T2A element at the same time. The map o...

Embodiment 2

[0065] The difference between this example and Example 1 is that the composite extract in step 3.1 is a mixture of digitonin, TritonX100, NP40, and CHAPS in a volume ratio of 1:1:1:1.

Embodiment 3

[0067] The difference between this example and Example 1 is that the composite extract in step 3.1 is a mixture of digitonin, TritonX100, NP40, and CHAPS in a volume ratio of 1:2:2:1.

[0068] When the anti-GABABR autoantibody detection material prepared in the above examples is used to detect the GABABR antibody in the sample, the present invention firstly prepares a new blocking material. Since some proteins in human serum and cerebrospinal fluid will non-specifically bind to antigens during the detection process and interfere with the detection results, the present invention uses a sealing material to seal the detection sample, and the implementation effect shows that the sample background can be significantly reduced, as follows A specific example of the preparation of the sealing material of the present invention is given.

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Abstract

The invention discloses detection materials, preparation methods and applications of anti-GABABR autoantibodies in human body fluids. In the invention, GABABR antigens are coated on NC membranes to make anti-GABABR receptor detection materials, and specific GABABR antibodies in human serum and cerebrospinal fluid It will bind to the antigen, use the alkaline phosphatase substrate-ligand reaction to develop color, and add the blocking material of the present invention in the color reaction, and can directly judge whether the detection sample contains GABABR antibody by visual observation. The method has the advantages of high sensitivity, simple operation, rapid detection, etc., and is helpful for the identification and diagnosis of anti-GABABR receptor encephalitis.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a detection material, preparation method and application of anti-GABABR autoantibody in human body fluid. Background technique [0002] Anti-gamma-aminobutyric acid receptor (GABABR) encephalitis belongs to neuroimmune encephalitis, most of which have epilepsy as the first symptom and symptoms of limbic encephalitis such as memory disturbance and psychosis, about 50 Paraneoplastic neoplasms occur in 100% of patients, most of which are related to small cell lung cancer, and the prognosis is poor. Anti-GABABR encephalitis was first reported by Lancaster et al. in 2010. Using immunoprecipitation and immunoblotting methods, 15 patients with anti-GABABR encephalitis were detected in the blood or cerebrospinal fluid of 410 patients with suspected paraneoplastic encephalitis or autoimmune encephalitis. GABABR antibody. Romana et al. detected 20 cases of GABABR antibodi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/705C07K1/14C12N15/867G01N33/531G01N33/564
CPCC07K14/70571C12N15/86C12N2740/15043C12N2800/107G01N33/531G01N33/564G01N2333/70571G01N2800/28
Inventor 闫亚平黎培李科
Owner SHAANXI MYBIOTECH CO LTD
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