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Application of a signal amplification technology in pgp9.5 detection kit

A detection kit and detection reagent technology, applied in the biological field, can solve the problems of low sensitivity, narrow linearity, long detection time, etc., and achieve the effect of reducing misjudgments and missed judgments

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

AI Technical Summary

Problems solved by technology

However, this invention focuses on protecting proteins, and related methods have long detection time, low sensitivity and narrow linearity

Method used

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  • Application of a signal amplification technology in pgp9.5 detection kit
  • Application of a signal amplification technology in pgp9.5 detection kit
  • Application of a signal amplification technology in pgp9.5 detection kit

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0084] Antibody 1 and antibody 2 are both self-made monoclonal antibodies; the above-mentioned preparation method of monoclonal antibody includes the following steps:

[0085] 1. Immune animals

[0086] BALB / c mice were selected as host animals for immunization, and the antigen solution of 1-5 mg / mL (the antigen was PGP9.5 protein, and the sequence of the PGP9.5 protein was shown in SEQ ID NO: 1) Intraperitoneal injection was performed after mixing with Freund's complete adjuvant at a volume ratio of 1:1. On the 14th and 35th days, the antigen solution of 1-5 mg / mL was mixed with incomplete Freund's adjuvant at a volume ratio of 1:1, and the second and third immunizations were carried out. The fourth immunization was performed with 5 mg / mL antigen in PBS. Cell fusion can be performed around day 61. During this period, two titer tests were performed on the 21st day and the 42nd day respectively to observe the immune effect.

[0087] Freund's complete adjuvant: sigma, F5881 ...

Embodiment 1

[0120] (1) Preparation of buffer solution

[0121] Buffer 1: Weigh 14.8g of ethanolamine and 5.8g of NaCl into a certain amount of purified water and stir until completely dissolved, adjust the pH value between 7.3 and 7.6 and set the volume to 1000ml. Filtration was performed with a 0.22 μm filter.

[0122] Buffer 2: Weigh 75g of glycine, add it to a certain amount of purified water, stir until completely dissolved, and make up to 1000ml. Filtration was performed with a 0.22 μm filter.

[0123] Buffer 3: Weigh 12.0g of Tris, 20.0g of bovine serum albumin, 2.0g of NaCl, and 10.0g of sucrose, add them to a certain amount of purified water and stir until completely dissolved, adjust the pH value between 7.5 and 10.0 and set the volume to 1000ml. Filtration was performed with a 0.22 μm filter.

[0124] Buffer 4: Weigh 12.0g of Tris, 9.0g of NaCl, 1.0g of bovine serum albumin, 5.0g of glycerol, and 5.0g of glycine, add them to a certain amount of purified water and stir until ...

Embodiment 2

[0171] (1) Preparation of buffer solution

[0172] Buffer 1: Weigh 15.1g of ethanolamine and 6.0g of NaCl into a certain amount of purified water and stir until completely dissolved, adjust the pH value between 7.3 and 7.6 and set the volume to 1000ml. Filtration was performed with a 0.22 μm filter.

[0173] Buffer 2: Weigh 75g of glycine, add it to a certain amount of purified water, stir until completely dissolved, and make up to 1000ml. Filtration was performed with a 0.22 μm filter.

[0174] Buffer 3: Weigh 15.0g of Tris, 60g of bovine serum albumin, 10.0g of NaCl, and 100g of sucrose into a certain amount of purified water and stir until completely dissolved, adjust the pH value between 7.5 and 10.0 and set the volume to 1000ml. Filtration was performed with a 0.22 μm filter.

[0175] Buffer 4: Weigh 15.0g of Tris, 9.0g of NaCl, 50g of bovine serum albumin, 20.0g of glycerol, and 40g of glycine into a certain amount of purified water and stir until completely dissolved...

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Abstract

The invention provides an application of a signal amplification technology in a PGP9.5 detection kit, and relates to the field of biotechnology. The kit includes a detection reagent strip; wherein, the detection reagent strip includes a reagent A, a reagent B and a reagent C; the The raw material of reagent A includes PGP9.5 antibody 1; the raw material of reagent B includes PGP9.5 antibody 2; the reagent C includes anti-FITC antibody. The present invention improves on the basis of the original invention, adds a "FITC-anti-FITC antibody" signal amplification system, increases the sensitivity of the kit, reduces the usage amount of main raw materials, and reduces the material cost.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of a signal amplification technology in a PGP9.5 detection kit. Background technique [0002] Traumatic brain injury (TBI) is brain damage caused by external forces that disrupts normal brain function, resulting in impaired cognitive or physical functioning. Among all types of TBI, the most common sequelae were headache (47.9%) and memory abnormalities (42%) (British Journal of Neurosurgery, 2018), and about 30% of patients required psychological counseling or neurological treatment. TBI is the most common disease in neurosurgery and one of the major causes of death and disability worldwide, and its sequelae have a permanent impact on the health of patients. [0003] The medical care process for suspected TBI is divided into three steps, starting with neurological assessment using the 15-point Glasgow Coma Scale (GCS) (American College of Surgeons Trauma Committee, 1...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/573G01N33/58G01N33/543G01N33/577G01N33/535G01N33/533C07K16/40
CPCG01N33/573G01N33/581G01N33/582G01N33/54326G01N33/577G01N33/535G01N33/533C07K16/40G01N2333/948G01N2800/28C07K2317/56C07K2317/92
Inventor 王法龙李锋杨涛孙佳张广俊张明琛
Owner SOPHONIX CO LTD
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