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Method and reagent for measuring thyroglobulin

A technique for thyroglobulin and immunoassay, which is used in biological testing, preparation of samples for testing, material testing, etc. to reduce the effect of interaction

Pending Publication Date: 2021-11-09
FUJIREBIO CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The method described in Patent Document 1 is effective, but it is necessary to use substances resistant to acidifying agents and surfactants as antibodies used in immunoassays

Method used

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  • Method and reagent for measuring thyroglobulin
  • Method and reagent for measuring thyroglobulin
  • Method and reagent for measuring thyroglobulin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0060]

[0061] (1) Preparation of anti-thyroglobulin antibody plate

[0062] Into a polystyrene 96-well microplate (Thermo F16 Black Maxisorp), 100 μL / well was injected with an antibody dilution solution (0.1M sodium bicarbonate, 0.1M sodium chloride, pH 9.6), and incubated overnight at 4°C. The microplate was washed three times with PBS, and then, 350 μL / well of blocking solution (1.0% BSA, 3% sucrose, PBS) was injected into each well, and incubated at room temperature for 3 hours. After removing the blocking solution, the plate was vacuum-dried to prepare an anti-Tg antibody plate.

[0063] (2) Production of alkaline phosphatase-labeled mouse anti-human Tg monoclonal antibody

[0064] Mouse anti-human Tg antibody 5E6 (manufactured by AbD Serotec) was enzyme-labeled according to a conventional method to prepare an alkaline phosphatase-labeled mouse anti-human Tg monoclonal antibody (labeled antibody). Antibody diluent (0.02M KH 2 PO 4 , 0.076M K 2 PO 4 , 0.25M NaCl,...

Embodiment 2

[0074]

[0075] For the autoantibody-positive model sample of Example 1, various surfactants (nonionic surfactants (Triton X-100 , Brij35), zwitterionic surfactant (CHAPS, C12APS, C14APS), anionic surfactant (SDS, NLS)) or denaturant (urea), in addition, by the method identical with embodiment 1, The measurement of Tg was carried out. During the measurement under each condition, the sample pretreated with the pretreatment liquid not containing surfactant and denaturant was also measured simultaneously. The measured counts under each condition are shown in Table 2.

[0076] 【table 2-1】

[0077]

[0078] 【Table 2-2】

[0079]

[0080] 【Table 2-3】

[0081]

[0082] 【Table 2-4】

[0083]

[0084] 【Table 2-5】

[0085]

[0086] 【Table 2-6】

[0087]

[0088] 【Table 2-7】

[0089]

[0090] 【Table 2-8】

[0091]

[0092] Table 2 shows that, in addition to alkali treatment, adding a nonionic surfactant, zwitterionic surfactant, anionic surfactant, or urea ...

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Abstract

Disclosed are a method and a reagent for measuring thyroglobulin, whereby it becomes possible to measure the amount of thyroglobulin more accurately by a single test without being affected by the interference with an anti-thyroglobulin antibody, and it also becomes possible to use an antibody having low resistance to acidifying agents and surfactants in immunoassays. A method for measuring thyroglobulin in a sample isolated from a living body by an immunoassay comprises a pretreatment step of mixing the sample isolated from the living body with a pretreatment solution containing an alkaline substance.

Description

technical field [0001] The invention relates to a method for measuring thyroglobulin and a reagent for measuring it. Background technique [0002] Thyroglobulin (Tg) is a glycoprotein with a molecular weight of 660,000 produced only by thyroid follicular cells. Biosynthesized Tg is released into the follicular lumen. In this process, through the action of peroxidase, the iodine molecule combines with the tyrosine group in the Tg molecule to synthesize thyroid hormone. The Tg in the follicular lumen is taken up again by the follicular cells and decomposed in the follicular cells, causing the release of thyroid hormone. Furthermore, this process is activated through the action of thyroid hormone (TSH). Therefore, in normal times, the release of Tg itself in the blood occurs only slightly, and the release of Tg in the blood indicates some abnormality of the thyroid gland. Therefore, Tg is a highly organ-specific and extremely useful marker for thyroid diseases. In particul...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/48G01N33/53
CPCG01N33/78G01N1/28G01N33/53
Inventor 大植千春八木慎太郎青柳克己
Owner FUJIREBIO CO LTD
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