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Method for labeling OxLDL-Ab by utilizing chloramine T

A chloramine and labeling technology, which is applied in the preparation methods of peptides, chemical instruments and methods, preparations for in vivo tests, etc., can solve the problems of poor stability, poor repetition rate, and long labeling time, and achieve good repeatability. Effect

Pending Publication Date: 2017-06-20
CHINA INSTITUTE OF ATOMIC ENERGY
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

In 2009, Liu Zihua and others published a report in the magazine "Isotope" entitled " 125 I-labeled oxidized low-density lipoprotein antibody "article, the article discloses the method of labeling OxLDL-Ab by the Iodogen iodination method, which has defects such as long labeling time (>8min), poor stability, and poor repetition rate, so it is urgently needed Development of a new method for labeling OxLDL-Ab with iodine

Method used

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  • Method for labeling OxLDL-Ab by utilizing chloramine T

Examples

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Embodiment 1

[0033] A method utilizing chloramine T labeling OxLDL-Ab, the method comprises the following steps:

[0034](1) Add PBS buffer solution and NaI solution in sequence to the reaction vessel and mix them evenly, then add OxLDL-Ab solution into the mixed solution with a pipette, and when adding, the lower end of the pipette should be extended to the mixing point. 2mm inside the liquid surface of the solution, mix again evenly; then add chloramine T, and cover the bottle cap within 2 seconds, the reaction time is 1min;

[0035] The PBS buffer is a phosphate buffer with a concentration of 0.2mg / ml and a pH value of 7.3; the NaI is Na 123 I; the concentration of chloramine T solution is 5mg / ml; OxLDL-Ab solution and Na 123 The dosage ratio of the I solution is 30 μg / 28MBq; the reaction temperature is room temperature.

[0036] (2) in the reaction product that step (1) obtains, add the Na of sodium metabisulfite 5mg / ml 2 S 2 o 5 solution to terminate the reaction, Na 2 S 2 o 5...

Embodiment 2

[0049] The difference from Example 1 is that the OxLDL-Ab is an IgM type OxLDL-Ab with a molecular weight of 900KDa, exists as a pentamer, and is connected by a disulfide bond. In step (1), use the pipette solution to add OxLDL-Ab solution, and the lower end of the pipette will extend into the 1mm place in the liquid surface of the mixed solution when adding, and the NaI is NaI 125 1; the concentration of the chloramine T solution is 2mg / ml, the concentration of the PBS buffer solution is 0.1mg / ml, and the pH value is 7.0; the consumption ratio of the OxLDL-Ab solution and the NaI solution is 35 μg / 28MBq; the reaction time is 2min .

[0050] Na in step (2) 2 S 2 o 5 The concentration of the solution is 2.5mg / ml; Na 2 S 2 o 5 The molar ratio with chloramine T is 1.2;

[0051] The volume ratio of bovine serum albumin in PBS buffer solution in step (3) is 2%.

Embodiment 3

[0053] Different from Example 1, the pipette adds the OxLDL-Ab solution to the mixed solution, and the lower end of the pipette will extend to 3mm in the liquid surface of the mixed solution when adding; the NaI is NaI 131 1; the concentration of the chloramine T solution is 6mg / ml, the concentration of the PBS damping fluid is 0.4mg / ml, and the pH value is 7.8; the consumption ratio of OxLDL-Ab solution and NaI solution is 40 μ g / 28MBq; The reaction time is 5min.

[0054] Na in step (2) 2 S 2 o 5 The concentration of the solution is 6mg / ml; Na 2 S 2 o 5 The molar ratio with chloramine T is 2.4;

[0055] The volume ratio of serum protein in PBS buffer solution in step (3) is 3%.

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Abstract

The invention belongs to the technical field of drug labeling and discloses a method for labeling OxLDL-Ab by utilizing chloramine T. The method comprises the following steps: (1) sequentially adding a PBS buffer, a NaI solution, an OxLDL-Ab solution and chloramine T in a reaction vessel, covering a bottle cap within 2 to 10 seconds and reacting for 1 to 5 minutes; (2) adding a sodium metabisulfite Na2S2O5 solution to the resultant reaction product so that the reaction is stopped, wherein the concentration of Na2S2O5 solution is 2.5 to 6 mg / ml, and the molar ratio of Na2S2O5 to chloramine T is 1.2 to 2.4; (3) leaching a PD-10 column with the PBS buffer until column equilibrium; (4) allowing the resultant reaction product obtained from the step (2) to pass the equilibrated PD-10 column obtained from the step (3). The method has the beneficial effects of high labeling rate, short labeling time, good stability, high radiochemical purity, and easiness in standardized operation.

Description

technical field [0001] The invention belongs to the technical field of drug labeling, and in particular relates to a method for labeling OxLDL-Ab with chloramine T. Background technique [0002] OxLDL-Ab is a kind of antibody that uses OxLDL as an antigen to induce the body's own immune response, and the OxLDL-Ab produced for different antigenic determinants is also different. At present, it is recognized in the academic circle that oxidized low-density lipoprotein (OxLDL) plays a key role in the formation and development of atherosclerosis, and is the most important marker in the early stage and the whole course of atherosclerosis. [0003] The close correlation between OxLDL-Ab and AS (atheroma) has also been recognized, and the in vitro accurate quantitative detection of important factors such as OxLDL-Ab in biological samples has been realized. Usually OxLDL-Ab is of IgG type, with a molecular weight ranging from a few kDa to tens of kDa (Da is Dalton). At present, Chen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C07K1/13A61K51/10A61K101/02
CPCA61K51/1018C07K16/18
Inventor 罗志福徐燕华李帅刘子华陈英李凤林樊彩云邓新荣
Owner CHINA INSTITUTE OF ATOMIC ENERGY
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