Method for marking Cys-Annexin V by use of 99mTc and application of method

A 99mtc-cys-annexinv and 99mtc technology, which is applied in peptide preparation methods, chemical instruments and methods, animal/human peptides, etc., can solve the problem of difficult separation of labeled products and radioactive impurities, and achieve high exchange labeling efficiency and separation Good effect and simple reaction process

Active Publication Date: 2013-06-19
JIANGSU INST OF NUCLEAR MEDICINE +1
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  • Abstract
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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 method with simple steps, high labeling rate and no need for further separation 99m Method for Tc labeling Cys-Annexin V;

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  • Method for marking Cys-Annexin V by use of 99mTc and application of method
  • Method for marking Cys-Annexin V by use of 99mTc and application of method
  • Method for marking Cys-Annexin V by use of 99mTc and application of method

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Embodiment 1

[0043] by 99m Tc-labeled Cys-Annexin V

[0044] Cys-Annexin V was provided by Jiangsu Target Biomedical Research Institute Co., Ltd. Using genetic engineering technology to clone, express and purify in Escherichia coli, add a cysteine ​​to the C-terminus of Annexin V, purify with metal affinity chromatography, and recombine Cys-Annexin V. See Chinese patent CN102690345A for specific preparation steps and features.

[0045] Cys-Annexin V by ligand exchange method 99m Tc mark, specific steps include:

[0046] (1) Take 100μL of 5mg / mL sodium ethylenediaminetetraacetic acid (EDTA-2Na) phosphate buffer (0.1M, pH7.4), and add 50μL of 1mg / mL Cys-Annexin V phosphate buffer (0.1M , pH7.4) and 15 μL 1mg / mL stannous chloride dilute hydrochloric acid (1% hydrochloric acid) for reaction;

[0047] (2) Take 0.5mL freshly rinsed Na 99m TCO 4 Solution (provided by the Nuclear Medicine Department of the Clinical Department of the Jiangsu Institute of Atomic Medicine), with a radiation do...

Embodiment 2

[0050] Carry out marking process according to the method described in embodiment 1, and its difference is only in order to 99m Tc-Cys-Annexin V is distinguished from other radioactive impurities, Cys-Annexin V is not added during the labeling process, and other operations remain unchanged, the obtained reaction products are mainly 99m Tc-EDTA, carry out HPLC detection with the detection condition described in the step (3) in embodiment 1, such as image 3 as shown, 99m The retention time of Tc-EDTA (t R ) is 13.4min.

Embodiment 3

[0052] Carry out marking process according to the method described in embodiment 1, and its difference is only in order to 99m Tc-Cys-Annexin V is distinguished from other radioactive impurities, Cys-Annexin V and sodium edetate are not added during the labeling process, while other operations remain unchanged, the obtained reaction products are mainly 99m Tc colloid, carry out HPLC detection with the detection condition described in the step (3) in the embodiment 1, such as Figure 4 as shown, 99m The retention time of Tc colloid (t R ) is 12.5min.

[0053] Carry out HPLC detection with the detection condition described in the step (3) in the embodiment 1, Na 99m TCO 4 Solution retention time (t R ) is 15.9min, such as Figure 5 shown.

[0054] Figure 1-5 Description in 99m Tc -Cys-Annexin V and other radioactive impurities (such as 99m Tc colloid, 99m Tc-EDTA and Na 99m TCO 4 ) can be completely separated, calculated by HPLC 99m The labeling rate of Tc -Cys-An...

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Abstract

The invention belongs to the technical field of SPECT (single photon emission computed tomography) development, and particularly relates to a method for marking Cys-Annexin V by use of 99mTc, and an application of 99mTc-Cys-Annexin V in detecting cell apoptosis as a developer. The method for marking Cys-Annexin V by use of 99mTc provided by the invention realizes a 99mTc marking technology for Cys-Annexin V through a ligand exchange method, and adopts ethylene diamine tetraacetate as a ligand in the ligand exchange method; the whole reaction has remarkably high exchange efficiency, and the marked product can be obviously distinguished from possible impurities in reaction; and the method has obvious advantages compared with a marking method of the prior art.

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 A method for Tc labeling Cys-Annexin V, and 99m Application of Tc-Cys-Annexin V as an imaging agent in the detection of 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 ph...

Claims

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Application Information

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Patent Type & Authority Applications(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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