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Blood platelet marker and preparation method thereof

A platelet and marker technology, applied in the field of biomedicine, can solve problems such as difficulty in directly and effectively detecting platelet aggregation site and degree of aggregation, lack of platelets, stagnation of research on thrombotic diseases, etc., and achieves excellent labeling effect, simple preparation method, The effect of good application prospects

Inactive Publication Date: 2014-03-26
LUZHOU MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, there has been a lack of platelet markers for a long time, making it difficult to directly and effectively detect the aggregation site and degree of platelet aggregation in clinical practice, making the research on thrombotic diseases stagnant. Therefore, it is urgent to find a platelet marker

Method used

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  • Blood platelet marker and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Preparation of platelet markers of the present invention

[0026] (1) Synthesis of 2G ~ 6G PAMAM:

[0027] Dissolve 1.4g of PAMAM in 70ml of methanol, add 100ul of sodium methoxide, and add 3ml of methyl acrylate dropwise under nitrogen protection. Stir for 72 hours, evaporate methanol and excess methyl acrylate to obtain a yellow oily liquid, which is 2G ~ 6G PAMAM.

[0028] (2) Combination of 2G-6G PAMAM with anti-CD41 monoclonal antibody:

[0029] Take 40ul of 2G-6G PAMAM prepared in step (1), add sulfo-SMCC40ul (4.8mg / mL) and anti-CD41 monoclonal antibody 20ul (200μg / ml) at room temperature, react for 2 hours, and pass the remaining reagents through gel filtration After 2 hours, add N-ethylmaleimide 40ul (4.8mg / mL) and react for 1 hour to obtain a reaction mixture; purify by ultrafiltration (MWCO100,000), wash with PBS, and obtain 2G-6G of anti-CD41 monoclonal antibody PAMAM.

[0030] (3) Take 20 μL of 2-6GPAMAM linked with anti-CD41 monoclonal antibod...

Embodiment 2

[0031] Example 2 Preparation of platelet markers of the present invention

[0032] (1) Synthesis of 2G ~ 6G PAMAM:

[0033] Dissolve 1.4g of PAMAM in 56ml of methanol, add 84ul of sodium methoxide, and add 2.8ml of methyl acrylate dropwise under nitrogen protection. Stir for 60 hours, evaporate methanol and excess methyl acrylate to obtain a yellow oily liquid, which is 2G ~ 6G PAMAM.

[0034] (2) Combination of 2G-6G PAMAM with anti-CD41 monoclonal antibody:

[0035] Take 30ul of 2G-6G PAMAM prepared in step (1), add 40ul of sulfo-SMCC (4.5mg / mL) and 20ul of anti-CD41 monoclonal antibody (150μg / ml) at room temperature, react for 1.5 hours, and pass the remaining reagents through gel filtration After 2 hours, N-ethylmaleimide 40ul (5.0mg / mL) was added and reacted for 0.5 hours to obtain a reaction mixture; purified by ultrafiltration (MWCO100,000) and washed with PBS, the 2G~ 6G PAMAM.

[0036] (3) Take 20 μL of 2G-6G PAMAM linked with anti-CD41 monoclonal antibody obtaine...

Embodiment 3

[0037] Example 3 Preparation of platelet markers of the present invention

[0038] (1) Synthesis of 2G ~ 6G PAMAM:

[0039] Dissolve 1.4g of PAMAM in 84ml of methanol, add 112ul of sodium methoxide, and add 3.22ml of methyl acrylate dropwise under nitrogen protection. Stir for 60 hours, evaporate methanol and excess methyl acrylate to obtain a yellow oily liquid, which is 2G ~ 6G PAMAM.

[0040] (2) Combination of 2G-6G PAMAM with anti-CD41 monoclonal antibody:

[0041] Take 50ul of 2G-6G PAMAM prepared in step (1), add sulfo-SMCC40ul (5.0mg / mL) and anti-CD41 monoclonal antibody 20ul (250μg / ml) at room temperature, react for 2.5 hours, and pass the remaining reagents through gel filtration After 2 hours, add N-ethylmaleimide 40ul (5.0mg / mL) and react for 2 hours to obtain a reaction mixture; purify by ultrafiltration (MWCO100,000), wash with PBS, and obtain 2G-6G with anti-CD41 monoclonal antibody PAMAM.

[0042] (3) Take 20 μL of 2G-6G PAMAM linked with anti-CD41 monoclon...

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Abstract

The invention discloses a blood platelet marker which is 2 to 6G PAMAM connected with a blood platelet specific antibody and fluorescent molecules. The invention also provides a preparation method of the blood platelet marker. The blood platelet marker disclosed by the invention is simple in preparation method, can effectively mark blood platelets and has a wide application prospect.

Description

technical field [0001] The invention relates to a platelet marker and a preparation method thereof, belonging to the technical field of biomedicine. Background technique [0002] Platelets are one of the key components of thrombus formation in vivo, and play an important role in various pathophysiological processes such as cardiovascular diseases (proliferation, thrombosis, and thromboembolism), inflammation, and some tumors. [0003] However, there has been a lack of platelet markers for a long time, which has made it difficult to directly and effectively detect the aggregation site and degree of platelets clinically, making the research on thrombotic diseases stagnant. Therefore, it is urgent to find a platelet marker. Contents of the invention [0004] In order to solve the above problems, the present invention provides a platelet marker and a preparation method thereof. [0005] The platelet marker of the present invention is 2-6G PAMAM connected with platelet-specifi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/533
CPCG01N33/533G01N33/582G01N2800/222G01N2800/32
Inventor 吴剑波罗茂李蓉
Owner LUZHOU MEDICAL COLLEGE
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