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Immunological measurement reagent for use in measurement of kl-6

An immunoassay, KL-6 technology, applied in the field of immunoassay reagents for KL-6 assay, can solve the problem that non-specific reaction samples cannot be sufficiently suppressed and the like

Active Publication Date: 2012-04-18
SEKISUI MEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, the present inventors have found that samples in which non-specific reactions occur cannot be sufficiently suppressed even by the above method

Method used

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  • Immunological measurement reagent for use in measurement of kl-6
  • Immunological measurement reagent for use in measurement of kl-6
  • Immunological measurement reagent for use in measurement of kl-6

Examples

Experimental program
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preparation example Construction

[0064]

[0065] Briefly, an aqueous solution of 1100 g of distilled water, 200 g of styrene, 0.2 g of sodium styrene sulfonate, and 1.5 g of potassium persulfate dissolved in 50 g of distilled water was prepared in an Mix in a 2-L glass reaction vessel. The atmosphere of the vessel was replaced with nitrogen, and the mixture was polymerized at 70° C. for 48 hours with stirring.

[0066] After polymerization, the above solution was filtered through filter paper and latex particles were recovered. The diameter of the obtained latex particles was measured by image analysis of at least 100 particles. Photographs for latex particle image analysis were taken at 10,000× magnification using a transmission electron microscope (JEM-1010, JEOL). The average diameter is 0.2 μm.

Embodiment 1

[0069] (Preparation of solution containing latex particles immobilized with anti-KL-6 antibody (solution containing latex particles immobilized thereon with anti-KL-6 antibody; hereinafter referred to as reagent 2))

[0070] The anti-KL-6 antibody solution (5mM Tris-HCl, pH 8.0) adjusted to 0.7 mg / mL was added to an equivalent amount of a solution (5mM Tris-HCl, pH 8.0) containing 1.0% latex particles having an average particle size of 0.2 μm. 0). After stirring at 4°C for 2 hours, an equal amount of 2.0% BSA solution (5 mM Tris-HCl, pH 8.0) was added, and the mixture was stirred at 4°C for 1 hour. After centrifugation, the supernatant was discarded, and the pellet was resuspended in 5 mM Tris-HCl, pH 8.0. The mixture was diluted with 5 mM Tris-HCl, pH 8.0, so that the absorbance at a wavelength of 600 nm was 4.5 Abs. This solution was used as reagent 2.

[0071] Preparation of a solution containing a rheumatoid factor interference inhibitor (hereinafter, reagent 1).

[00...

Embodiment 2 to 4

[0080] Instead of 30 mM citrate buffer (pH 4.0) in Reagent 1 of Example 1, 30 mM citrate buffer at pH 4.5 (Example 2), pH 5.0 (Example 3) and pH 5.5 (Example 4) was used . Measurements were performed using the same reagents and the same assay as in Example 1.

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Abstract

Disclosed are a measurement reagent and a measurement method both for accurately measuring KL-6, particularly a measurement reagent and a measurement method for accurately measuring KL-6 in a sample that contains a rheumatoid factor and / or a non-specific substance other than the rheumatoid factor. Specifically disclosed is an immunological measurement reagent which comprises a solution containing a rheumatoid factor interference inhibitor and having a pH value of 4.0 to 5.5 and a solution containing an insoluble carrier, wherein an anti-KL-6 antibody is supported on the insoluble carrier. The use of the immunological measurement reagent enables the accurate measurement of KL-6 in a sample that contains a rheumatoid factor and / or a non-specific substance other than the rheumatoid factor.

Description

technical field [0001] The technical field of the invention relates to assay reagents and assays for the measurement of KL-6 in samples. Furthermore, the technical field of the present invention relates to immunoagglutination techniques, reagents and methods of inhibiting non-specific reactions in this technique. Background technique [0002] KL-6 is a sialylated carbohydrate antigen involved in pulmonary fibrosis (Non-Patent Document (NPL) 1, Patent Document (PTL) 1). Since elevated KL-6 levels and their fluctuations in interstitial pneumonia indicate pathological signs (PTL 1), measurement of KL-6 levels is used in the diagnosis of interstitial pneumonia and determination of treatment strategies. A method of predicting the onset of interstitial pneumonia caused by interferon administration by measuring serum MUC-1 / KL-6 levels has been disclosed (PTL 2), a method of examining prognosis in lung cancer patients by measuring KL-6 (PTL 3 ), and a method for detecting intraduc...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/543
CPCG01N2333/4725G01N33/6884Y10S436/805Y10S436/808Y10S436/815G01N33/543
Inventor 近藤纯一山本光章金子智惠
Owner SEKISUI MEDICAL CO LTD
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