Oncofetal fibronectin detection method using simple immunoassay

A fibronectin and carcinoembryonic technology, applied in the field of detection of carcinoembryonic fibronectin using immunochromatography, can solve the problems of not describing the similarity of antigens, etc., and achieve the effect of reducing cross-reactivity, preventing measurement errors or false positive reactions

Inactive Publication Date: 2018-11-09
SEKISUI MEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0014] Furthermore, neither of Patent Documents 1 and 2 describes or suggests problems related to the antigenic similarity between fFN and other fibronectins

Method used

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  • Oncofetal fibronectin detection method using simple immunoassay
  • Oncofetal fibronectin detection method using simple immunoassay
  • Oncofetal fibronectin detection method using simple immunoassay

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0183] [Example 1] Relationship between the type of surfactant added to the sample extract and FN cross-reactivity

[0184] The type of surfactant added to the sample extract was changed by using the detection device fabricated above, and its influence on the cross-reactivity of plasma FN was confirmed. For concentrations of nonionic surfactants, % means % (w / v) unless otherwise indicated.

[0185] 1. Test method

[0186] (1) Sample preparation

[0187] The normal human plasma pool was diluted with the fFN sample extract described below to prepare a 0.3% plasma sample.

[0188]

[0189] As shown in Table 1, 10 mM PBS, 1% BSA, 1 mM EDTA-2Na, nonionic surfactant (0.1%, 0.5%).

[0190] [Table 1]

[0191]

nonionic surfactant

1

TritonX-114 (prior art)

2

Triton X-100

3

n-Heptyl-β-D-thioglucoside

4

n-Octyl-β-D-glucoside

5

Tween 20

6

Brij35 (Brij is a trademark)

[0192] (2) Measurement

[0193] 120 ...

Embodiment 2

[0197] [Example 2] Relationship between the added concentration of Brij 35 and FN cross-reactivity

[0198] 1. Test method

[0199] Detect in the same manner as in Example 1, except that the normal human plasma confluence (pool) is treated with 0.2%, 0.4%, 0.5%, 0.6% or 0.8% Brij 35 as a nonionic surfactant Dilute the fFN sample extract to adjust the plasma concentration to 0.2% and 0.5%. As a comparative example, the fFN sample extract (prior art) added with Triton X-114 was also tested.

[0200] 2. Test results

[0201] The results are shown in figure 2 middle.

[0202] Compared with the addition of 0.1% Triton X-114 in the prior art, when Brij 35 was added in the concentration range of 0.4% to 0.8%, the cross-reactivity with plasma FN was inhibited.

Embodiment 3

[0203] [Example 3] Relationship between Brij chain length and FN cross-reactivity

[0204] 1. Test method

[0205] Detect in the same manner as in Example 1, except that the plasma confluence of normal people is diluted with the fFN sample extract containing 0.1% or 0.5% of the nonionic surfactants shown in Table 2, to Plasma concentrations were adjusted to 0.2% and 0.3%. As a comparative example, the fFN sample extract (prior art) added with 0.1% or 0.5% Triton X-114 was also tested.

[0206] [Table 2]

[0207]

[0208] 2. Test results

[0209] The results are shown in image 3 middle.

[0210] Both concentrations of Brij 35 (m=23) and Brij O20 (m=20) inhibited the cross-reactivity with plasma fibronectin, thereby exhibiting a high inhibitory effect, compared to the same concentration of Triton X-114. In contrast, 0.5% Brij S100 (m = 100) and Brij 93 (m = 2) could not inhibit the cross-reactivity with plasma fibronectin compared with the same concentration of Triton ...

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Abstract

The present invention addresses the problem of making it possible, in a simple immunoassay, to prevent false-positive reactions and reduce measurement values errors so as to accurately detect fFN by suppressing a cross-reaction with plasma FN that is an antigenic component similar to fFN to reduce the detection of plasma FN. When an anti-fFN antibody reacts with fFN in the presence of a specific surfactant according to the present invention, it is possible to accurately detect fFN through the suppression of the cross-reaction between the antibody and FN, even when other FNs such as plasma FN are also present. The specific surfactant is represented by general formula (1). In the formula, m is 4-30, and R represents an alkyl group.R-O-(CH2Ch2O)mH ...(1).

Description

technical field [0001] The present disclosure relates to a method for detecting carcinoembryonic fibronectin using a simple immunoassay, and in particular to a method for detecting carcinoembryonic fibronectin using immunochromatography. Furthermore, the present invention relates to a method capable of accurately detecting carcinoembryonic fibronectin in a case where fibronectin other than carcinoembryonic fibronectin may coexist in a sample. Background technique [0002] Point of Care Testing (POCT) is an umbrella term for testing usually performed in the vicinity of a patient, such as a general practitioner, a hospital ward, or a clinic for outpatient treatment. Since diagnosis and treatment can be performed quickly according to test results obtained by simple operations, it is expected to contribute to the improvement of the quality of medical treatment. [0003] The detection method utilizing immunochromatography is a commonly used method in POCT, which is used in pract...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/543
CPCG01N2333/78G01N33/689G01N33/5306G01N33/558G01N33/57476G01N33/54387G01N33/54393G01N33/543G01N33/574G01N33/54388
Inventor 伊藤佳菜子森田元喜家治翔平酒井骏
Owner SEKISUI MEDICAL CO LTD
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