Immunoassay method and assay reagent used in said method

A technology for antibody detection and samples, applied in biological testing, measuring devices, material inspection products, etc., can solve problems such as the difference between the measured value and the real value, interference hindering the detection of antigens and antibodies, and damage to the accuracy of the measured value

Inactive Publication Date: 2018-01-02
SEKISUI MEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although immunological assay methods are characterized by high specificity and good sensitivity, various interferences are also known to hinder the detection of specific reactions between antigens and antibodies
The non-specific reaction due to interference destroys the accuracy of the measured value and causes the problem that the obtained measured value is different from the real value

Method used

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  • Immunoassay method and assay reagent used in said method
  • Immunoassay method and assay reagent used in said method
  • Immunoassay method and assay reagent used in said method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0179] [Example 1] Confirmation of non-specific reaction inhibitory effect of BPF on samples with L-FABP concentration near normal value, part 1

[0180] The non-specific reaction inhibitory effect in the L-FABP concentration range around the normal value when no protein (Comparative Example 1), BSA (Comparative Example 1) or 0.1% BPF (Example 1) was added to the first reagent was compared.

[0181] 1. Operation

[0182] (1) Embodiment 1

[0183] Using frozen-thawed urine as a sample, L-FABP in the sample was determined by using a first control reagent containing 390 mmol / L of the compound of formula (1) (benzamidine hydrochloride) and 0.1% BPF and a second reagent.

Embodiment 2 to 5

[0193] [Examples 2 to 5] Confirmation of non-specific reaction inhibitory effect of BPF on samples having L-FABP concentrations near normal values, part 2

[0194] Compare 390mmol / L compound (benzamidine hydrochloride) of formula (1) and 0.01% to 1% BPF (embodiment 2 to 5) when being added to the nonspecific in the L-FABP concentration range near the normal value when the first reagent Heterosexual response suppression effect.

[0195] 1. Operation

[0196] For the samples, a portion of urine (n=10; measured value in the reference method 0.6 ng / mL to 4.9 ng / mL) frozen at -30°C after collection was thawed only once and used for measurement. The amount of BPF indicated in Table 2 was added to the first reagent. Example 2 was performed in the same manner as Example 1 except these points.

[0197] 2. Results

[0198] The net absorbances of the samples measured in the experiments of Examples 2 to 5 were converted into L-FABP concentrations by using a calibration curve establish...

Embodiment 6 to 9

[0202] [Examples 6 to 9] Confirmation of non-specific reaction inhibitory effect of BPF on samples having L-FABP concentrations near normal values, part 3

[0203] Compare 390mmol / L compound (2-amino-2-thiazoline hydrochloride) of formula (2) and 0.01% to 1% BPF (embodiments 6 to 9) when adding to the first reagent near the normal value L-FABP Inhibitory effect of non-specific reactions in the concentration range.

[0204] 1. Operation

[0205] For the samples, a portion of urine (n=10; measured value in the reference method 0.6 ng / mL to 4.9 ng / mL) frozen at -30°C after collection was thawed only once and used for measurement. The compound of formula (2) (2-amino-2-thiazoline hydrochloride) and the amount of BPF shown in Table 3 were added to the first reagent. Example 3 was performed in the same manner as Example 1 except these points.

[0206] 2. Results

[0207] The net absorbances of the samples measured in the experiments of Examples 6 to 9 were converted into L-FABP ...

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Abstract

The purpose of the present invention is to reduce the possibility of nonspecific reactions occurring during the immunoassay of L-FABPs, and provide an assay method that is more accurate and has good sensitivity. More specifically, the purpose of the present invention is to provide an assay method that is sensitive and accurate, even when the concentration of a substance to be assayed is low (for example, when the concentration of L-FABPs is near normal levels). A L-FABP immunoassay method using anti-L-FABP antibodies, wherein it is possible to easily suppress nonspecific reactions even duringsensitive assays by including a polypeptide comprising the amino acid sequence represented by SEQ ID NO: 1, which is the sequence from the 419th amino acid to the 607th amino acid in the amino acid sequence of the heat shock protein (HSP) DnaK from Escherichia coli, or by including a polypeptide having a sequence identity of at least 90% therewith.

Description

technical field [0001] The present invention relates to an immunological assay method for L-FABP using an anti-L-FABP antibody and an assay reagent used in the method. Background technique [0002] Fatty acid binding proteins (FABP) are a group of proteins that exist in the cytosol and have a molecular weight of about 14 kilodaltons and the ability to bind fatty acids, and include at least seven known molecular species, such as the liver type (L-FABP) , intestinal type (I-FABP), heart type (H-FABP), brain type (B-FABP), skin type (C-FABP / E-FABP), adipocyte type (aP2) and peripheral nerve cell type (myeloid Phospholipids P2), these families are thought to have evolved from a common ancestral gene. Although each type of FABP shows a specific tissue distribution, the name indicates the tissue in which the type was first found, not necessarily that the type is present only in that tissue. For example, at least two types of FABP, liver type (L-FABP) and heart type (H-FABP), are...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/531G01N33/53G01N33/543
CPCG01N33/54313G01N33/54393G01N33/6893G01N33/53G01N2333/47
Inventor 小林幸司松本拓二山本光章
Owner SEKISUI MEDICAL CO LTD
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