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Cys-Annexin V kit used for 99mTc labeling and preparation method and application thereof

A 99mtc, medicine box technology, applied in the field of single photon emission computed tomography (SPECT) imaging, can solve the problem of difficult separation of labeling products and radioactive impurities, and achieve the effects of intuitive and accurate labeling results, accurate results and simple operation.

Active Publication Date: 2015-07-01
JIANGSU INST OF NUCLEAR MEDICINE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] For this reason, the technical problem to be solved by the present invention lies in the 99m In the method of Tc labeling Cys-Annexin V, it is difficult to separate the labeling product from radioactive impurities, and then provide a simple step, high labeling rate, which can be used for 99m Kits for Tc-labeled Cys-Annexin V variants;

Method used

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  • Cys-Annexin V kit used for 99mTc labeling and preparation method and application thereof
  • Cys-Annexin V kit used for 99mTc labeling and preparation method and application thereof
  • Cys-Annexin V kit used for 99mTc labeling and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0049] The specific preparation process of the kit described in this example is as follows: Weigh 1.9 mmol of sodium dihydrogen phosphate and 8.1 mmol of disodium hydrogen phosphate, dissolve them in 90 mL of distilled water, add 150 mg of sodium edetate, and add 2.00 mg / mL of Add 1mL of stannous chloride dilute hydrochloric acid (1% hydrochloric acid) solution, and finally add 1.5mL of 5mg / mL Cys-Annexin V solution, mix well, filter aseptically, and then divide into 10mL vials at 1.00mL / bottle. Put the vial into the freeze dryer, set the temperature of the freeze dryer to -40°C, freeze for 4 hours, then turn on the vacuum pump to vacuum (after the vacuum pump is turned on, the whole process is completed in a vacuum state until the medicine box Freeze-drying is complete), make the vacuum pressure reach 1-2Pa, then freeze for 2 hours, adjust the temperature to -20°C, freeze for 3 hours, then adjust the temperature to -10°C, freeze for 13 hours, then adjust the temperature to 10°...

Embodiment 2

[0053] by 99m Method for Tc labeling Cys-Annexin V kit

[0054] Get the kit prepared in Example 1, add 1ml of newly rinsed Na 99m TCO 4 solution, placed in a 35-37°C water bath for 15 minutes to obtain the labeled product 99m Tc-Cys-Annexin V, the Na 99m TCO 4 The solution was provided by the Nuclear Medicine Department of the Clinical Department of Jiangsu Institute of Atomic Medicine, and the radiation dose was 18.5-555MBq.

[0055] Use HPLC method to detect the labeled product, use TSK GEL gel column (SWG2000swxL), use phosphate buffer solution with concentration 0.05M, pH 7.0 as mobile phase, flow rate 0.8mL / min, UV detection wavelength 278nm, and set There is an isotope detector. At the same time, unlabeled Cys-Annexin V was detected with the same detection conditions. Such as Figure 1-2 shown, measured 99m The retention time of Tc-Cys-Annexin V (t R ) was 9.8min, consistent with the retention time of unlabeled Cys-Annexin V. Through HPLC chromatogram analysis...

Embodiment 3

[0057] Prepare the kit according to the method described in Example 1, the difference is only in order to 99m Tc-Cys-Annexin V is distinguished from other radioactive impurities, and Cys-Annexin V is not added when preparing the kit, so that the resulting kit does not contain the labeled target Cys-Annexin V, while other preparation and labeling operations unchanged, the obtained labeled products are mainly 99m Tc-EDTA, carry out HPLC detection with the detection condition of HPLC purification among the embodiment 2, as image 3 as shown, 99m The retention time of Tc-EDTA (t R ) is 13.4min.

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Abstract

The invention belongs to the development technical field of single photon emission computed tomography (SPECT) and in particular relates to a Cys-Annexin V kit used for 99mTc labeling, a preparation method and application thereof in apoptosis detection. Each kit provided by the invention is prepared as follows: 25-150ug of Cys-Annexin V, 0.5-6mg of edetate, 5-40ug of tin salt and 0.1mmol of pH buffer agent. When the kit provided by the invention carries out 99mTc labeling for Cys-Annexin V, the operation is simple, the labeling rate is stable and larger than 95%, and the labeled product can be obviously distinguished from the impurities possibly appearing during the reaction. Compared with the labeling method in the existing technology, the method provided by the invention has obvious advantages.

Description

technical field [0001] The invention belongs to the technical field of single photon emission computed tomography (SPECT) imaging, and specifically relates to a 99m Tc-labeled Cys-Annexin V kit, its preparation method and its application in detecting cell apoptosis. Background technique [0002] Apoptosis, also known as programmed cell death (PCD), is an important part of the cell life cycle and is an active cell death process regulated by genes that is different from necrosis and accidental death. Apoptosis involves a series of changes in biomolecules and cell morphology. Among them, phosphatidylserine (PS) in the lipid bilayer of the cell membrane is flipped from the inner layer to the outer layer and exposed to the cell surface, which is an early event of apoptosis. , occurs before the morphological changes of apoptotic cells, and is also the initial event of the apoptotic cascade reaction, as a sign that apoptotic cells are recognized by phagocytes, which further causes...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/47C07K1/13A61K51/08A61K103/10
Inventor 陆春雄蒋泉福俞惠新华子春胡敏进季宇
Owner JIANGSU INST OF NUCLEAR MEDICINE
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