Protein for binding tafia/ai and application thereof and kit for detecting tafia/ai content

A kit and protein technology, applied in the biological field, can solve the problems of complicated operation of detection methods, unstable detection results, low detection sensitivity, etc., and achieve the effects of improving detection sensitivity, low detection limit and low cost.

Active Publication Date: 2020-10-30
LIAONING MEDI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The first object of the present invention is to provide a protein for binding TAFIa / ai, the second object of the present invention is to provide the application of the above protein in binding TAFIa / ai, the third object of the present invention is to provide a A kind of kit for detecting TAFIa / ai content, to alleviate the detection method existing in the prior art for the possibility of occurrence of cardiovascular and cerebrovascular diseases etc. The operation is complicated, the equipment is expensive, the detection sensitivity is low, the detection result is unstable, and the detection time is relatively long. long technical question

Method used

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  • Protein for binding tafia/ai and application thereof and kit for detecting tafia/ai content
  • Protein for binding tafia/ai and application thereof and kit for detecting tafia/ai content
  • Protein for binding tafia/ai and application thereof and kit for detecting tafia/ai content

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] The preparation of embodiment 1 TAFIa / ai chemiluminescent quantitative detection reagent

[0059] (1) Construction of PGT1 protein

[0060] Entrust a technical service company to synthesize the PGT1 protein sequence designed in the present invention according to the codon preference of Escherichia coli BL21. The protein sequence is as follows (SEQ ID NO.1):

[0061] EQHADPICNKPCKTHDDCSGAWFCQACWNSARTCGPYVGMKLYFSRNPNPRLAVAIARYLETKLDFEFASPFAAGQMEKFRRLNPNLSLPILVDDEGKSLWEADAIACRLSRHAHSDFWRTGDDEPEMIRWLSWGKEHFALACDTVHFERGTKQRYGIGPIDQKRVEEGLNQFHTAAAMLDAVLAERQWLVGNSVSYADFRMATFLPFNDAARLPLDDYPSVSRWYRRLEDIDAWRDPFKGMDAPELPPVPQMAVADQREQYDPVCHKPCSTQDDCSGGTFCQACWRFAGTCGPYVHHHHHH。

[0062] The physical map of the obtained vector is as figure 1 shown.

[0063] The prepared protein was subjected to electrophoresis, and the results were as follows: figure 2 Shown: each lane is ①purified PGT1 protein; ②purified PGT1 protein; ③purified PGT1 protein; ④unpurified PGT1 protein; M protein ma...

Embodiment 2

[0090] Example 2 TAFIa / ai Chemiluminescent Quantitative Detection Method I

[0091] (1) The model of the chemiluminescence instrument is BPCL weak luminescence measuring instrument. The detection cell temperature was set at 37°C.

[0092] (2) Add 150 μL of the reagent I-a provided in step (6) of Example 1 of the present invention, 150 μL of reagent I-b provided in step (9) of Example 1 of the present invention and 5 μL of the standard in the detection tube, mix After homogenization, place in the detection pool and incubate for 10 min.

[0093] (3) Magnetic separation, remove the supernatant. Inject 50 μL each of liquid A and liquid B in the chemiluminescent substrate I provided in step (11) of Example 1 of the present invention into the detection tube in sequence, and start recording the luminescence intensity for 300 s. In this experiment, the photon count corresponding to the peak area of ​​the chemiluminescence kinetic curve was collected.

[0094] (4) Establish a stand...

Embodiment 3

[0098] Example 3 TAFIa / ai Chemiluminescent Quantitative Detection Method II

[0099] (1) The model of the chemiluminescence instrument is BPCL weak luminescence measuring instrument. The detection cell temperature was set at 37°C.

[0100] (2) Add 150 μL of the reagent II-a provided in step (7) of Example 1 of the present invention, 150 μL of reagent II-b provided in step (10) of Example 1 of the present invention and 5 μL of the standard in the detection tube, mix After homogenization, place in the detection pool and incubate for 10 min.

[0101] (3) Magnetic separation, removing the supernatant, and adding 300 μL of the chemiluminescent cleaning solution provided in step (13) of Example 1 of the present invention. Remove the magnetic field, mix well and incubate in the detection cell for 5 minutes.

[0102] (4) Inject the chemiluminescent substrate II provided in step (12) of Example 1 of the present invention into the detection tube, start recording the luminescence inte...

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PUM

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Abstract

The invention provides a protein for being combined with TAFIa / ai and an application thereof, and a kit for detection of the TAFIa / ai content, and relates to the biotechnology field; the protein for being combined with the TAFIa / ai has the amino acid sequence as shown in SEQ ID NO.1. The protein can be specifically combined with the TAFIa / ai and is characterized in that each protein molecule can be combined with two TAFIa / ais at the same time, and the detection sensitivity is improved. The kit for detecting the content of the TAFIa / ai includes the protein or a TAFIa / ai antibody. The kit has the advantages of simple and fast operation, low cost, sensitive diagnosis, low detection limit and the like. As a new TAFIa / ai detection method, the kit can give further help to clinical thrombus disease research and TAFIa / ai detection.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a protein for binding TAFIa / ai and its application and a kit for detecting the content of TAFIa / ai. Background technique [0002] As we all know, thrombotic disease is a kind of disease that seriously affects health, especially cardiovascular and cerebrovascular embolism disease has become the first cause of death in our country. Due to the variety of clinical manifestations of thrombotic diseases, their clinical diagnosis and treatment are often very complicated. At present, clinical examination methods for thrombotic diseases mainly include electrocardiography, echocardiography, chest X-ray, blood pressure monitoring, cranial X-ray, brain CT scan, brain MRI examination, DSA, MRA, transcranial Doppler ultrasonography, etc. Radiographic examinations will produce a certain degree of ionizing radiation to the human body. There are many functional imaging examination methods for tissue...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/00G01N33/68G01N33/573
Inventor 李文欣苏春阳刘峰
Owner LIAONING MEDI BIOTECH CO LTD
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