WO2018016670A1 - Biochip for diagnosing medicinal herb allergy - Google Patents

Biochip for diagnosing medicinal herb allergy Download PDF

Info

Publication number
WO2018016670A1
WO2018016670A1 PCT/KR2016/008376 KR2016008376W WO2018016670A1 WO 2018016670 A1 WO2018016670 A1 WO 2018016670A1 KR 2016008376 W KR2016008376 W KR 2016008376W WO 2018016670 A1 WO2018016670 A1 WO 2018016670A1
Authority
WO
WIPO (PCT)
Prior art keywords
biochip
immunoglobulin
allergy
herbal medicine
strip
Prior art date
Application number
PCT/KR2016/008376
Other languages
French (fr)
Korean (ko)
Inventor
황제환
김범준
임국진
최윤아
김태형
이지은
최동섭
Original Assignee
(주)프로테옴텍
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by (주)프로테옴텍 filed Critical (주)프로테옴텍
Publication of WO2018016670A1 publication Critical patent/WO2018016670A1/en

Links

Images

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6854—Immunoglobulins
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54386—Analytical elements
    • G01N33/54387—Immunochromatographic test strips
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
    • G01N2333/705—Assays involving receptors, cell surface antigens or cell surface determinants
    • G01N2333/70503—Immunoglobulin superfamily, e.g. VCAMs, PECAM, LFA-3
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/24—Immunology or allergic disorders

Definitions

  • the present invention relates to a biochip for diagnosing medicinal herbs allergy, and more particularly, to a biochip for diagnosing allergy caused by herbal medicine by measuring the concentration of immunoglobulin (antibody, antibody) in a biological sample reacting with the herbal medicine.
  • Biochip is a substrate made in the form of a chip by combining materials such as DNA, proteins, enzymes, antibodies, microorganisms, animal and plant cells, organs, and neurons derived from living organisms to a solid substrate made of an inorganic material. It is a device that can analyze biological information such as gene expression pattern, gene binding, disease diagnosis, protein distribution using large scale.
  • Biochips that measure proteins are commonly used in autoimmune antibodies and allergy diagnostic reagents.
  • Conventional biochip kits provide diagnostic test results through measurement of substances in biological samples for various diseases or biological immune causing agents.
  • existing inventions do not consider herbal medicines widely used in East Asia as candidate agents.
  • IgE immunoglobulins
  • herbal medicine means a herbal medicine that is taken from an animal, plant or mineral and is mainly dried, cut or purified in a circular shape.
  • herbal medicines were safe and had no side effects, but the recent reports on the side effects of herbal medicines have increased the demand for stability of taking herbal medicines.
  • allergies caused by herbal medicines are widely known because of their frequent dryness.
  • the current situation is not dealing with allergic reactions caused by other herbs.
  • the present inventors have tried to develop a biological material measurement tool and method for herbal medicine.
  • a biological material measurement tool and method for herbal medicine As a result, by measuring the concentration of specific immunoglobulins (antibodies, antibodies) for the herbal medicines possessed by individual patients, a method for in vitro diagnosis of allergic reactions to the herbal medicines was identified.
  • an object of the present invention is to solve the above problems, and to measure the concentration of specific immunoglobulin (antibody, antibody) in a biological sample reacting with the herbal medicine for diagnosing allergy caused by herbal medicine for diagnosing allergy caused by herbal medicine It is to provide a biochip.
  • Still another object of the present invention is to provide a method for diagnosing allergy of herbal medicines by measuring a concentration of specific immunoglobulins (antibodies, antibodies) present by contacting a biological sample to the diagnostic biochip marker of the above term.
  • the present invention provides a herbal medicine allergy diagnosis biochip for diagnosing allergy caused by herbal medicine by measuring the concentration of specific immunoglobulin (antibody, antibody) in a biological sample reacting with the herbal medicine.
  • the biochip comprises a strip in one reaction vessel; And a container for fixing the strip; It may be to include.
  • the strip may be selected from the group consisting of line type, dot type, parallel line type.
  • the strip may include a herb allergen (arllergen) as a marker.
  • a herb allergen arllergen
  • the medicinal herb allergen is root, licorice, dried, antler, baekrye, daily donkey, earth, creation, cheongung, pedestrians, Astragalus, Jimo, ginseng, fennel, shiho , Motherwort, raw grass, gardenia, leaf, peony, baekryeok, ferry, fermentation, peony, golden, bee venom, changja, wolfberry, cornus, golboji, hyunsam, wood, wooden , Seokchangpo, Seshin, Schisandra, Taxa, may be one or more of the group consisting of McMoon-Dong.
  • the strip may be to coat a membrane (membrane) on the support.
  • the membrane may be selected from the group consisting of nitrocellulose, nylon, polyvinylidene fluoride (PVDF), glass and plastic.
  • PVDF polyvinylidene fluoride
  • the present invention provides a herbal medicine allergy diagnosis method for measuring the concentration of a specific immunoglobulin (antibody, antibody) present in the sample by contacting the biological sample to the diagnostic biochip marker.
  • the immunoglobulin may include one or more of the group consisting of immunoglobulin E (Immunoglobulin E, IgE) or immunoglobulin G (Immunoglobulin G, IgG).
  • the biological sample may be selected from the group consisting of tissues, cells, whole blood, blood serum, plasma, saliva, cerebrospinal fluid, and urine.
  • the present invention has been made to solve the above problems, the present invention is to diagnose the allergic reaction caused by the herbal medicine through the measurement results of specific immunoglobulins (antibodies, antibodies) present in the biological sample using the herbal medicine allergen as a marker To provide a biochip for.
  • the present invention is to provide a method for diagnosing allergy caused by herbal medicine by contacting a biological sample to the biochip marker, by measuring the concentration of immunoglobulin (antibody, antibody).
  • Figure 1 shows a schematic diagram of the line-type biochip of the present invention.
  • Figure 2 shows a schematic diagram of the dot-type biochip of the present invention.
  • Figure 3 shows a schematic diagram of a parallel line-type biochip of the present invention.
  • Figure 4 is a biochip subjected to a reaction process according to Example 2 of the present invention by contacting the herbal strip prepared in parallel line form with a biological sample.
  • Example 5 is a biochip subjected to a reaction process according to Example 3 of the present invention by contacting a herbal strip prepared in a parallel line form with a biological sample.
  • the present invention sought to solve the above-mentioned problem by finding an allergic reaction diagnostic biochip caused by herbal medicine through the measurement results of specific immunoglobulins (antibodies, antibodies) present in a biological sample using the herbal medicine allergen as a marker.
  • allergic reactions to herbal medicines can be known indirectly through in vitro diagnosis, and it is effective to provide information for taking herbal medicines more safely by preventing side effects in advance.
  • the present invention provides a herbal medicine allergy diagnosis biochip for diagnosing allergy caused by herbal medicine by measuring the concentration of specific immunoglobulin (antibody, antibody) in a biological sample reacting with the herbal medicine.
  • the biochip of the present invention comprises a strip in one reaction vessel; And a container for fixing the strip; Characterized in that it comprises a.
  • the strip 10 is disposed in one biochip and includes a container 20 to hold the strip (see FIGS. 1 to 3).
  • the strip may be selected from the group consisting of line, dot, parallel line.
  • the 'line strip' refers to a strip prepared by attaching a marker in a line form on the membrane, and the term 'drawing a line' means attaching a marker on the membrane.
  • the 'dot-type strip' refers to a strip prepared by attaching a marker in the form of dots on a membrane, and the meaning of 'dotting' means to attach a marker on the membrane.
  • the line strip is mainly because the markers to be measured on the liver membrane are drawn in the form of strips, each of which is lined up horizontally, and then cut vertically.
  • the dot strip can be used for the detection of various materials, and the parallel line strip can detect a wide range of materials with only a small amount of sample, and the time and manpower required for detection can be significantly reduced. There is an advantage to production.
  • the strip may include a herb allergen as a marker.
  • Allergens or allergens are allergens that cause allergic reactions, in which allergens produce specific immunoglobulin (antibody) antibodies against specific foreign substances.
  • allergens produce specific immunoglobulin (antibody) antibodies against specific foreign substances.
  • allergen diagnosis it is very important to simultaneously diagnose various allergens that can cause allergies. Therefore, using the biochip of the present invention will have the effect that can be diagnosed simultaneously for a number of herbal allergens.
  • the herb allergens are black root, licorice, dry, antler, baekchul, perilla, earth, creation, cheonggung, passerby, yellow, gimmo, ginseng, fennel, shiho, motherwort, raw sulfur, gardenia, hubak, hwangbaekcho, bokyeong, jisil, Peony, golden, bee venom, changja, wolfberry, cornus, bogorji, hyeonsam, keg, doin, bridge, fast, mine, spirit, ancient bones, white paper, pesticide, seokchangpo, sesin, omija, taeksa, mammundong, etc. It is not limited.
  • the strip comprises a support.
  • the membrane is coated on the support, and the membrane may be nitrocellulose, nylon, polyvinylidene fluoride (PVDF), glass, plastic, or the like, but is not limited thereto.
  • PVDF polyvinylidene fluoride
  • the present invention also provides a herbal allergy diagnosis method using the biochip of the present invention.
  • Specific immunoglobulins (antibodies, antibodies) present in the biological sample can be detected by reaction with the marker, and the immunoglobulin immunoglobulin E (Immunoglobulin E, IgE) or immunoglobulin G (Immunoglobulin G, IgG)
  • the diagnosis of allergy is to measure the concentration level of a specific immunoglobulin (antibody, antibody) detected by comparing it with a control.
  • the biological sample may be, but is not limited to, tissue, cells, whole blood, blood serum, plasma, saliva, cerebrospinal fluid, or urine.
  • immunoglobulin is known to play an important role in the functional immunity of serum components, structural features having a characteristic or common tertiary structure in x-ray crystalography analysis, fractionated to a common location of electrophoresis It is a collective expression of a specific range of "glycoproteins” that have features that can be clearly distinguished from other types of proteins, in terms of physical characteristics to be synthesized, and features that have a certain common amino acid sequence when analyzing amino acid sequences.
  • immunoglobulin The basic structure of immunoglobulin is that a pair of L chains (light chains) having a molecular weight (dalton) of about 23,000 and a pair of H chains (heavy chains) having a molecular weight of about 50,000 to 70,000 are connected by SS bonds.
  • L chains light chains
  • H chains heavy chains
  • IgG IgA
  • IgM IgM
  • IgD IgD
  • IgE immunoglobulin
  • Specific concentration of the specific immunoglobulin (antibody, antibody) that is expressed may be used to measure the fluorescence, absorbance, magnetism, electrical flow and the like.
  • NC-membrane nitrocellulose membrane
  • sample diluent (0.5% BSA, PBS) was added to the membrane-coated plastic reaction container, and then 50 ⁇ l of serum of the herb allergy patient was added and stirred at room temperature for 45 minutes. After diluting the diluted serum solution in the reaction vessel, 400 ul of washing solution (50mM Tris, 0.2M NaCl, 0.05% Tween20) is aliquoted and stirred at room temperature for 5 minutes and then removed. This procedure was added once.
  • Example 2 Following Example 1 and Example 2, but using 300 ul of the Biotin-labeled mouse anti-phosphorus IgG antibody solution instead of 300 ul of the Biotin-labeled mouse anti-phosphorus IgE antibody solution of Example 2 was stirred for 30 minutes. The rest of the procedure was carried out in the same manner as in Example 1 and Example 2, and as a result, it was confirmed that the medicinal herb markers reacted with specific immunoglobulin G (s-IgG) were colored black. As in Example 2, the (-) strip is a sera test group of negative patients, and the (+) strip is a sera test group of positive patients. Identification and quantification of specific immunoglobulin G (s-IgG) is possible.
  • specific immunoglobulin G specific immunoglobulin G, IgG

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Chemical & Material Sciences (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

The present invention relates to a biochip for diagnosing a medicinal herb allergy which diagnoses an allergy caused by a medicinal herb by measuring the concentration of a specific immunoglobulin (antibody).

Description

한약재 알레르기 진단용 바이오칩Herbal Allergy Diagnosis Biochip
본 출원은 2016년 07월 18일 출원된 대한민국 특허출원 제10-2016-0091014호를 우선권으로 주장하고, 상기 명세서 전체는 본 출원의 참고문헌이다.This application claims the priority of Korean Patent Application No. 10-2016-0091014, filed on July 18, 2016, the entirety of which is a reference of the present application.
본 발명은 한약재 알레르기 진단용 바이오칩에 관한 것으로, 더욱 상세하게는 한약재와 반응하는 생체 시료 내 면역글로불린(항체, antibody)의 농도를 측정하여 한약재로 야기되는 알레르기를 진단하는 한약재 알레르기 진단용 바이오칩에 관한 것이다.The present invention relates to a biochip for diagnosing medicinal herbs allergy, and more particularly, to a biochip for diagnosing allergy caused by herbal medicine by measuring the concentration of immunoglobulin (antibody, antibody) in a biological sample reacting with the herbal medicine.
바이오칩(Biochip)은 무기물로 된 고체 기질에 생물에서 유래된 DNA, 단백질, 효소, 항체, 미생물, 동식물 세포 및 기관, 신경세포 등의 물질을 결합시켜 칩 형태로 만든 기판으로서 생체분자의 고유한 생화학적 특성을 이용하여 유전자 발현 양상, 유전자 결합, 질병의 진단, 단백질 분포 등의 생물학적 정보를 대규모로 분석할 수 있는 장치를 말한다. Biochip is a substrate made in the form of a chip by combining materials such as DNA, proteins, enzymes, antibodies, microorganisms, animal and plant cells, organs, and neurons derived from living organisms to a solid substrate made of an inorganic material. It is a device that can analyze biological information such as gene expression pattern, gene binding, disease diagnosis, protein distribution using large scale.
단백질을 측정하는 바이오칩은 자가 면역 항체, 알레르기 진단시약 등에서 많이 사용되고 있다. 종래의 바이오칩 키트들은 다양한 질병 또는 생체 면역반응 원인 물질에 대한 생체 시료 내 물질 측정을 통하여 진단 검사 결과를 제공한다. 그러나 기존의 발명들은 동아시아에서 널리 사용되는 한약재는 원인 후보 물질로 고려하고 있지 않다.Biochips that measure proteins are commonly used in autoimmune antibodies and allergy diagnostic reagents. Conventional biochip kits provide diagnostic test results through measurement of substances in biological samples for various diseases or biological immune causing agents. However, existing inventions do not consider herbal medicines widely used in East Asia as candidate agents.
혈액 내 특이 면역글로불린(항체, antibody)의 측정은 알레르기 질환 및 기생충 감염과 연관되어 있으며 거의 항상 알레르기 혈액 페널 검사의 테스트로 측정된다. 어떤 사람의 경우 대부분의 사람에서 해가 없고 면역 반응 또한 유발하지 않는 이물질에 대해서 면역 반응을 일으킬 수 있다. 이러한 반응을 알레르기(또는 과민성)이라고 하며, 이 반응은 IgE 항체의 생성을 수반한다. 알레르기 반응은 약한 자극부터 생명을 위협하는 심한 반응에 이르기까지 증상이 다양하며, 이에 대한 대처는 항히스타민제나 휴대용 에피네프린 주사와 같은 약물을 사용할 수 있으나, 사전에 알레르기 반응 유발 요인을 찾아 접촉을 피하는 것이 가장 확실한 방법이다. 혈액 내 IgE 측정은 반응 유발 요인에 대한 정보를 제공한다는 점에서 중요성을 지닌다. Measurement of specific immunoglobulins (antibodies) in the blood is associated with allergic diseases and parasitic infections and is almost always measured by tests of allergic blood panel tests. In some people, most people can develop an immune response against a foreign substance that is harmless and does not trigger an immune response. This response is called allergy (or hypersensitivity), which involves the production of IgE antibodies. Allergic reactions can range from mild irritation to life-threatening severe reactions, and medications such as antihistamines or portable epinephrine injections can be used, but it is best to avoid contact with the triggers beforehand. It's a sure way. The measurement of IgE in the blood is important in that it provides information on response triggers.
약사법 총칙 제2조 5항에 따르면, '한약'이란 동물, 식물 또는 광물에서 채취된 것으로 주로 원형대로 건조, 절단 또는 정제된 생약을 말한다. 과거 한약재는 안전하고 부작용이 없다고 인식되었으나 최근 한약의 부작용에 관한 보도가 많아지면서 한약 복용의 안정성에 대한 요구가 많아지고 있다. 여러 형태의 부작용 중, 한약재로 인한 알레르기는 건칠(옻)으로 인해 발생하는 경우가 그 빈번함으로 인해 널리 알려져 있다. 하지만 다른 한약재에 의한 알레르기 반응에 대해서는 다루고 있지 못한 것이 현재의 상황이다. 다양한 한약재에 대한 알레르기 반응을 확인하기 위한 검사법이 필요한 실정이지만 이를 위한 진단 도구나 기준이 전무한 상황이다.According to Article 2, Paragraph 5 of the General Regulations of the Pharmaceutical Affairs Act, "herbal medicine" means a herbal medicine that is taken from an animal, plant or mineral and is mainly dried, cut or purified in a circular shape. In the past, it was recognized that herbal medicines were safe and had no side effects, but the recent reports on the side effects of herbal medicines have increased the demand for stability of taking herbal medicines. Among various forms of side effects, allergies caused by herbal medicines are widely known because of their frequent dryness. However, the current situation is not dealing with allergic reactions caused by other herbs. Although there is a need for a test method to confirm allergic reactions to various herbal medicines, there are no diagnostic tools or criteria for this.
이에 본 발명자들은 한약재에 대한 생물학적 물질 측정도구 및 방법을 개발하고자 노력하였다. 그 결과, 개별 환자가 지닌 한약재에 대한 특이 면역글로불린(항체, antibody)의 농도를 측정함으로써, 한약재에 대한 알레르기 반응을 체외 진단하는 방법을 규명하였다.Therefore, the present inventors have tried to develop a biological material measurement tool and method for herbal medicine. As a result, by measuring the concentration of specific immunoglobulins (antibodies, antibodies) for the herbal medicines possessed by individual patients, a method for in vitro diagnosis of allergic reactions to the herbal medicines was identified.
따라서, 본 발명의 목적은 상기와 같은 문제점을 해결하기 위해 안출된 것으로, 한약재와 반응하는 생체 시료 내 특이 면역글로불린(항체, antibody)의 농도를 측정하여 한약재로 야기되는 알레르기를 진단하는 한약재 알레르기 진단용 바이오칩을 제공하는 것이다.Accordingly, an object of the present invention is to solve the above problems, and to measure the concentration of specific immunoglobulin (antibody, antibody) in a biological sample reacting with the herbal medicine for diagnosing allergy caused by herbal medicine for diagnosing allergy caused by herbal medicine It is to provide a biochip.
본 발명의 또 다른 목적은 상기 항의 진단용 바이오칩 마커에 생물학적 시료를 접촉시켜, 존재하는 특이 면역글로불린(항체, antibody)의 농도를 측정하는 한약재 알레르기 진단 방법을 제공하는 것이다. Still another object of the present invention is to provide a method for diagnosing allergy of herbal medicines by measuring a concentration of specific immunoglobulins (antibodies, antibodies) present by contacting a biological sample to the diagnostic biochip marker of the above term.
그러나, 본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 과제에 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당업자에게 명확하게 이해될 수 있을 것이다. However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problem, another task that is not mentioned will be clearly understood by those skilled in the art from the following description.
따라서, 본 발명은 한약재와 반응하는 생체 시료 내 특이 면역글로불린(항체, antibody)의 농도를 측정하여 한약재로 야기되는 알레르기를 진단하는 한약재 알레르기 진단용 바이오칩을 제공한다.Accordingly, the present invention provides a herbal medicine allergy diagnosis biochip for diagnosing allergy caused by herbal medicine by measuring the concentration of specific immunoglobulin (antibody, antibody) in a biological sample reacting with the herbal medicine.
상기 본 발명의 바람직한 일실시예에 따르면, 상기 바이오칩은 하나의 반응 용기 내에 스트립(strip); 및 상기 스트립을 고정하는 용기; 를 포함하는 것일 수 있다.According to a preferred embodiment of the present invention, the biochip comprises a strip in one reaction vessel; And a container for fixing the strip; It may be to include.
상기 본 발명의 바람직한 다른 일실시예에 따르면, 상기 스트립은 라인형, 도트형, 병렬식 라인형으로 이루어진 군으로부터 선택되는 것일 수 있다.According to another preferred embodiment of the present invention, the strip may be selected from the group consisting of line type, dot type, parallel line type.
상기 본 발명의 바람직한 또 다른 일실시예에 따르면, 상기 스트립은 한약재 알레르겐(arllergen)을 마커로 포함하는 것일 수 있다.According to another preferred embodiment of the present invention, the strip may include a herb allergen (arllergen) as a marker.
상기 본 발명의 바람직한 또 다른 일실시예에 따르면, 상기 한약재 알레르겐(allergen)은 갈근, 감초, 건칠, 녹용, 백출, 일당귀, 지구자, 창출, 천궁, 행인, 황기, 지모, 인삼, 회향, 시호, 익모초, 생지황, 치자, 후박, 황백초, 복령, 지실, 작약, 황금, 봉독, 창이자, 구기자, 산수유, 보골지, 현삼, 목통, 도인, 연교, 속단, 마인, 저령, 고본, 백지, 산약, 석창포, 세신, 오미자, 택사, 맥문동으로 이루어진 군 중 1종 이상 포함하는 것일 수 있다. According to another preferred embodiment of the present invention, the medicinal herb allergen (allergen) is root, licorice, dried, antler, baekrye, daily donkey, earth, creation, cheongung, pedestrians, Astragalus, Jimo, ginseng, fennel, shiho , Motherwort, raw grass, gardenia, leaf, peony, baekryeok, ferry, fermentation, peony, golden, bee venom, changja, wolfberry, cornus, golboji, hyunsam, wood, wooden , Seokchangpo, Seshin, Schisandra, Taxa, may be one or more of the group consisting of McMoon-Dong.
상기 본 발명의 바람직한 또 다른 일실시예에 따르면, 상기 스트립은 지지체 위에 멤브레인(membrane)을 코팅하는 것일 수 있다.According to another preferred embodiment of the present invention, the strip may be to coat a membrane (membrane) on the support.
상기 본 발명의 바람직한 또 다른 일실시예에 따르면, 상기 멤브레인은 니트로셀룰로스, 나일론, 폴리플루오린화비닐리덴(PVDF), 유리 및 플라스틱으로 이루어진 군으로부터 선택되는 것일 수 있다.According to another preferred embodiment of the present invention, the membrane may be selected from the group consisting of nitrocellulose, nylon, polyvinylidene fluoride (PVDF), glass and plastic.
또한, 본 발명은 상기 진단용 바이오칩 마커에 생물학적 시료를 접촉시켜, 시료 내에 존재하는 특이 면역글로불린(항체, antibody)의 농도를 측정하는 한약재 알레르기 진단 방법을 제공한다. In addition, the present invention provides a herbal medicine allergy diagnosis method for measuring the concentration of a specific immunoglobulin (antibody, antibody) present in the sample by contacting the biological sample to the diagnostic biochip marker.
상기 본 발명의 바람직한 일실시예에 따르면, 상기 면역글로불린은 면역글로불린 E(Immunoglobulin E, IgE) 또는 면역글로불린 G(Immunoglobulin G, IgG),로 이루어진 군 중 1종 이상 포함하는 것일 수 있다.According to a preferred embodiment of the present invention, the immunoglobulin may include one or more of the group consisting of immunoglobulin E (Immunoglobulin E, IgE) or immunoglobulin G (Immunoglobulin G, IgG).
상기 본 발명의 바람직한 다른 일실시예에 따르면, 상기 생물학적 시료는 조직, 세포, 전혈, 혈정, 혈장, 타액, 뇌척수액, 및 뇨로 이루어진 군으로부터 선택되는 것일 수 있다.According to another preferred embodiment of the present invention, the biological sample may be selected from the group consisting of tissues, cells, whole blood, blood serum, plasma, saliva, cerebrospinal fluid, and urine.
본 발명은 상기와 같은 문제점을 해결하기 위해 안출된 것으로, 본 발명은 한약재 알레르겐을 마커로 하여 생물학적 시료 내에 존재하는 특이 면역글로불린(항체, antibody)의 측정 결과를 통해 한약재로 인한 알레르기 반응을 진단하기 위한 바이오칩을 제공하고자 한다.The present invention has been made to solve the above problems, the present invention is to diagnose the allergic reaction caused by the herbal medicine through the measurement results of specific immunoglobulins (antibodies, antibodies) present in the biological sample using the herbal medicine allergen as a marker To provide a biochip for.
또한, 본 발명은 상기 바이오칩 마커에 생물학적 시료를 접촉시켜, 면역글로불린(항체, antibody)의 농도를 측정함으로써 한약재로 야기되는 알레르기 진단방법을 제공하고자 한다.In addition, the present invention is to provide a method for diagnosing allergy caused by herbal medicine by contacting a biological sample to the biochip marker, by measuring the concentration of immunoglobulin (antibody, antibody).
도 1는 본 발명의 라인형 바이오칩의 모식도를 나타낸 것이다.Figure 1 shows a schematic diagram of the line-type biochip of the present invention.
도 2는 본 발명의 dot형 바이오칩의 모식도를 나타낸 것이다.Figure 2 shows a schematic diagram of the dot-type biochip of the present invention.
도 3은 본 발명의 병렬식 라인형 바이오칩의 모식도를 나타낸 것이다.Figure 3 shows a schematic diagram of a parallel line-type biochip of the present invention.
도 4는 병렬식 라인형으로 제조된 한약재 스트립을 생물학적 시료와 접촉시켜 본 발명의 실시예 2에 따른 반응 과정을 거친 바이오칩이다.Figure 4 is a biochip subjected to a reaction process according to Example 2 of the present invention by contacting the herbal strip prepared in parallel line form with a biological sample.
도 5는 병렬식 라인형으로 제조된 한약재 스트립을 생물학적 시료와 접촉시켜 본 발명의 실시예 3에 따른 반응 과정을 거친 바이오칩이다.5 is a biochip subjected to a reaction process according to Example 3 of the present invention by contacting a herbal strip prepared in a parallel line form with a biological sample.
이하, 본 발명을 보다 상세하게 설명한다.Hereinafter, the present invention will be described in more detail.
상술한 바와 같이, 지금까지 질병을 진단하기 위한 바이오칩에 관한 보고가 있음에도 불구하고 한약재로 야기되는 알레르기를 진단하기 위한 바이오칩에 관한 연구는 없었다. As described above, there have been no studies on biochips for diagnosing allergies caused by herbal medicines, although there have been reports on biochips for diagnosing diseases.
본 발명은 한약재 알레르겐을 마커로 하여 생물학적 시료 내에 존재하는 특이 면역글로불린(항체, antibody)의 측정 결과를 통해 한약재로 인해 야기되는 알레르기 반응 진단 바이오칩을 밝혀냄으로써 상술한 문제의 해결을 모색하였다. 이를 통해 한약재에 대한 알레르기 반응을 체외 진단을 통해 간접적으로 알 수 있고, 이를 위한 부작용을 사전에 방지함으로써 한약을 더욱 안전하게 복용하기 위한 정보를 제공하는 효과가 있다. The present invention sought to solve the above-mentioned problem by finding an allergic reaction diagnostic biochip caused by herbal medicine through the measurement results of specific immunoglobulins (antibodies, antibodies) present in a biological sample using the herbal medicine allergen as a marker. Through this, allergic reactions to herbal medicines can be known indirectly through in vitro diagnosis, and it is effective to provide information for taking herbal medicines more safely by preventing side effects in advance.
본 발명은 한약재와 반응하는 생체 시료 내 특이 면역글로불린(항체, antibody)의 농도를 측정하여 한약재로 야기되는 알레르기를 진단하는 한약재 알레르기 진단용 바이오칩을 제공한다.The present invention provides a herbal medicine allergy diagnosis biochip for diagnosing allergy caused by herbal medicine by measuring the concentration of specific immunoglobulin (antibody, antibody) in a biological sample reacting with the herbal medicine.
본 발명의 바이오칩은 하나의 반응 용기 내에 스트립(strip); 및 상기 스트립을 고정하는 용기; 를 포함하는 것을 특징으로 한다. 스트립(10)이 하나의 바이오칩 안에 배치되어 있고, 상기 스트립을 고정할 수 있는 용기(20)를 포함하게 되는 것이다(도 1 내지 도3 참조)The biochip of the present invention comprises a strip in one reaction vessel; And a container for fixing the strip; Characterized in that it comprises a. The strip 10 is disposed in one biochip and includes a container 20 to hold the strip (see FIGS. 1 to 3).
상기 스트립은 라인형, 도트형, 병렬식 라인형으로 이루어진 군으로부터 선택되는 것일 수 있다. The strip may be selected from the group consisting of line, dot, parallel line.
본 발명에서 '라인형 스트립'은 멤브레인 위에 라인(line) 형태로 마커를 부착시켜 제조된 스트립을 의미하는 것으로, '라인을 긋는다'는 의미는 멤브레인 위에 마커를 부착시키는 것을 의미하는 것이다. 또한, 본 발명에서 '도트형 스트립'은 멤브레인 위에 도트(dot) 형태로 마커를 부착시켜 제조된 스트립을 의미하는 것으로 '도트를 찍는다'는 의미는 멤브레인 위에 마커를 부착시키는 것을 의미하는 것이다. 상기 라인형 스트립은 주로 간 멤브레인에 측정하고자 하는 마커들을 각각 라인 형태로 가로로 여러 줄을 그려놓고 세로로 잘라서 스트립 형태로 사용하기 때문에 생산 공정이 단순하고 다량 생산이 가능한 이점이 있다. 또한, 상기 도트형 스트립은 다양한 물질의 검출을 위해 사용될 수 있으며 상기 병렬식 라인형 스트립은 적은 시료만으로, 광범위한 물질을 검출할 수 있고 검출에 소모되는 시간 및 인력이 현저히 감소될 수 있으며 이를 통해 대량 생산이 가능한 이점이 있다. In the present invention, the 'line strip' refers to a strip prepared by attaching a marker in a line form on the membrane, and the term 'drawing a line' means attaching a marker on the membrane. In addition, in the present invention, the 'dot-type strip' refers to a strip prepared by attaching a marker in the form of dots on a membrane, and the meaning of 'dotting' means to attach a marker on the membrane. The line strip is mainly because the markers to be measured on the liver membrane are drawn in the form of strips, each of which is lined up horizontally, and then cut vertically. In addition, the dot strip can be used for the detection of various materials, and the parallel line strip can detect a wide range of materials with only a small amount of sample, and the time and manpower required for detection can be significantly reduced. There is an advantage to production.
본 발명에서 상기 스트립은 한약재 알레르겐을 마커로 포함하는 것일 수 있다. 알레르겐 또는 알레르기 항원은 알레르기를 유발하는 물질로서, 알레르기는 특정 외부 물질에 대한 특이 면역글로불린(항체, antibody) 항체가 체내에 생성되어 나타나는 과민성 면역반응이다. 이러한 알레르기 진단에 있어 알레르기를 일으킬 수 있는 다양한 알레르겐을 동시에 진단하는 것은 매우 중요하다. 따라서 본 발명의 바이오칩을 이용하면 다수의 한약재 알레르겐에 대하여 동시에 진단할 수 있는 효과를 가지게 되는 것이다.In the present invention, the strip may include a herb allergen as a marker. Allergens or allergens are allergens that cause allergic reactions, in which allergens produce specific immunoglobulin (antibody) antibodies against specific foreign substances. In allergy diagnosis, it is very important to simultaneously diagnose various allergens that can cause allergies. Therefore, using the biochip of the present invention will have the effect that can be diagnosed simultaneously for a number of herbal allergens.
상기 한약재 알레르겐은 갈근, 감초, 건칠, 녹용, 백출, 일당귀, 지구자, 창출, 천궁, 행인, 황기, 지모, 인삼, 회향, 시호, 익모초, 생지황, 치자, 후박, 황백초, 복령, 지실, 작약, 황금, 봉독, 창이자, 구기자, 산수유, 보골지, 현삼, 목통, 도인, 연교, 속단, 마인, 저령, 고본, 백지, 산약, 석창포, 세신, 오미자, 택사, 맥문동 등 일 수 있으나, 이에 제한되지 않는다.The herb allergens are black root, licorice, dry, antler, baekchul, perilla, earth, creation, cheonggung, passerby, yellow, gimmo, ginseng, fennel, shiho, motherwort, raw sulfur, gardenia, hubak, hwangbaekcho, bokyeong, jisil, Peony, golden, bee venom, changja, wolfberry, cornus, bogorji, hyeonsam, keg, doin, bridge, fast, mine, spirit, ancient bones, white paper, pesticide, seokchangpo, sesin, omija, taeksa, mammundong, etc. It is not limited.
본 발명의 일 구현예에 따르면, 상기 스트립은 지지체를 포함한다. 상기 지지체 위에는 멤브레인을 코팅하게 되며, 상기 멤브레인은 니트로셀룰로스, 나일론, 폴리플루오린화비닐리덴(PVDF), 유리, 또는 플라스틱 등 일 수 있으나, 마커를 포함할 수 있는 소재라면 이에 한정되는 것은 아니다. According to one embodiment of the invention, the strip comprises a support. The membrane is coated on the support, and the membrane may be nitrocellulose, nylon, polyvinylidene fluoride (PVDF), glass, plastic, or the like, but is not limited thereto.
또한, 본 발명의 바이오칩을 이용하는 한약재 알레르기 진단 방법을 제공한다. The present invention also provides a herbal allergy diagnosis method using the biochip of the present invention.
상기 생물학적 시료 내 존재하는 특이 면역글로불린(항체, antibody)는 상기 마커와의 반응을 통해 검출될 수 있고, 상기 면역글로불린 면역글로불린 E(Immunoglobulin E, IgE) 또는 면역글로불린 G(Immunoglobulin G, IgG)인 것일 수 있으며, 알레르기의 진단은 이를 대조군과 비교하여 검출되는 특이 면역글로불린(항체, antibody)의 농도 수준을 측정하는 것이다. 상기 생물학적 시료는 조직, 세포, 전혈, 혈정, 혈장, 타액, 뇌척수액, 또는 뇨 등일 수 있으나, 이에 제한되는 것은 아니다.Specific immunoglobulins (antibodies, antibodies) present in the biological sample can be detected by reaction with the marker, and the immunoglobulin immunoglobulin E (Immunoglobulin E, IgE) or immunoglobulin G (Immunoglobulin G, IgG) The diagnosis of allergy is to measure the concentration level of a specific immunoglobulin (antibody, antibody) detected by comparing it with a control. The biological sample may be, but is not limited to, tissue, cells, whole blood, blood serum, plasma, saliva, cerebrospinal fluid, or urine.
본 발명에서 "면역 글로불린"은 혈청성분 중 기능적으로 면역에 중요한 역할을 하는 것으로 알려져 있으며, x-ray crystalography 분석상 특징적인 또는 공통적인 3 차 구조를 가지는 구조적 특징, 전기영동상의 공통적인 위치로 분획화되는 물리적인 특징, 그리고 아미노산 서열 분석시 일정한 공통적인 아미노산 서열을 지니는 특징 등의 공통점으로 다른 종류의 단백질들과 명확히 구분할 수 있는 특징들을 지니는 특정 범위의 "당단백질"을 총칭하여 표현한 것이다. 면역글로불린은 기본구조는 분자량(dalton)이 약 2만 3000의 L사슬(경괘) 1쌍과 분자량 약 5만~7만의 H사슬(중쇄) 1쌍이 S-S 결합에 의해 연결되어 있으며, H 사슬의 종류인 γ, α, μ, δ, ε 에 의해 각각 IgG, IgA, IgM, IgD, IgE로 분류된다. 이 중 본 발명에서는 알레르기 반응과의 관련성이 규명된 IgE 및 IgG에 관해 다루고 있다.In the present invention, "immunoglobulin" is known to play an important role in the functional immunity of serum components, structural features having a characteristic or common tertiary structure in x-ray crystalography analysis, fractionated to a common location of electrophoresis It is a collective expression of a specific range of "glycoproteins" that have features that can be clearly distinguished from other types of proteins, in terms of physical characteristics to be synthesized, and features that have a certain common amino acid sequence when analyzing amino acid sequences. The basic structure of immunoglobulin is that a pair of L chains (light chains) having a molecular weight (dalton) of about 23,000 and a pair of H chains (heavy chains) having a molecular weight of about 50,000 to 70,000 are connected by SS bonds. Are classified into IgG, IgA, IgM, IgD, and IgE by γ, α, μ, δ, and ε. Among them, the present invention deals with IgE and IgG, which have been identified as being related to allergic reactions.
상기 발현되는 특이 면역글로불린(항체, antibody)의 농도 측정은 형광도, 흡광도, 자성, 전기흐름 등을 측정하는 방법이 사용될 수 있다. Specific concentration of the specific immunoglobulin (antibody, antibody) that is expressed may be used to measure the fluorescence, absorbance, magnetism, electrical flow and the like.
이하, 본 발명을 실시예에 의해 상세히 설명하기로 한다. 그러나 이들 실시예는 본 발명을 보다 구체적으로 설명하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail by way of examples. However, these examples are intended to illustrate the present invention in more detail, and the scope of the present invention is not limited to these examples.
실시예 1. 한약재 알레르겐 바이오칩의 제조Example 1 Preparation of Herbal Allergen Biochip
한약재들을 액체질소로 냉각시킨 후, 막자 사발을 이용하여 곱게 갈아주었다. 약 10 g의 한약재를 20 mM Tris(pH 8.5) 100 ml에 처리한 후 4에 밤새 교반을 통하여 한약재 알레르겐을 추출하였다. 이렇게 추출된 한약재 알레르겐 44종류를 나이트로셀룰로즈 멤브레인(NC-멤브레인)에 분주하였르며, 추출된 한약재 알레르겐의 종류를 하기 표 1에 나타냈다. 알레르겐이 고정된 NC-멤브레인은 Dry chamber에서 RT O/N건조하였다. 건조된 멤브레인을 플라스틱 지지체에 접착시키고 세로로 1.85 mm 간격으로 자른 후 직사각형의 플라스틱 반응용기에 양면 테이프를 이용하여 고정하였다. 멤브레인이 코팅된 플라스틱 반응 용기에 시료희석액(0.5% BSA, PBS) 250ul를 넣은 후 한약재 알레르기 환자의 혈청 50ul를 추가한 후 상온에서 45분동안 교반하였다. 반응 용기 안의 희석된 혈청액을 버린 후 세정액 (50mM 트리스, 0.2M NaCl, 0.05% Tween20) 400 ul를 분주하고 상온에서 5분간 교반한 후 제거한다. 이 과정을 1회 추가하였다.After cooling the herbal medicines with liquid nitrogen, finely ground using a mortar and pestle. About 10 g of the herbal medicine was treated with 100 ml of 20 mM Tris (pH 8.5), and then the herbal medicine allergen was extracted through stirring overnight at 4. Thus, 44 kinds of medicinal herb allergens were dispensed into nitrocellulose membrane (NC-membrane), and the types of the herbal medicinal allergens extracted are shown in Table 1 below. Allergen-fixed NC-membranes were dried in RT O / N in a dry chamber. The dried membrane was adhered to a plastic support, cut at 1.85 mm intervals, and fixed to a rectangular plastic reaction vessel using double-sided tape. 250 μl of sample diluent (0.5% BSA, PBS) was added to the membrane-coated plastic reaction container, and then 50 μl of serum of the herb allergy patient was added and stirred at room temperature for 45 minutes. After diluting the diluted serum solution in the reaction vessel, 400 ul of washing solution (50mM Tris, 0.2M NaCl, 0.05% Tween20) is aliquoted and stirred at room temperature for 5 minutes and then removed. This procedure was added once.
번호number 목록List 번호number 목록List
1One 갈근Rooting 2323 작약 Peony
22 감초 licorice 2424 황금 Gold
33 건칠 Dry 2525 봉독 Bee venom
44 녹용 velvet 2626 창이자 Changza
55 백출 Whiteness 2727 구기자 Wolfberry
66 일당귀One ear 2828 산수유 Cornus
77 지구자 Earth 2929 보골지 Golgol
88 창출 Creation 3030 현삼 figwort
99 천궁 Cheongung 3131 목통 Neck
1010 행인 Passerby 3232 도인 Doin
1111 황기 Astragalus 3333 연교 Fellowship
1212 지모 Jimo 3434 속단 Fast
1313 인삼 Ginseng 3535 마인 Mine
1414 회향 fennel 3636 저령 Bad
1515 시호 Shiho 3737 고본 Book
1616 익모초 motherwort 3838 백지blank
1717 생지황 Living 3939 산약 Medicine
1818 치자 Gardenia 4040 석창포 Seokchangpo
1919 후박 Thick 4141 세신 Sesin
2020 황백초Yellow white 4242 오미자 Schisandra
2121 복령 Ghost 4343 택사 Taxi
2222 지실 Basement 4444 맥문동Macmundong
실시예 2. 혈청 내 IgE 농도 측정Example 2. Determination of IgE Concentration in Serum
Biotin 표지 마우스 항-인 IgE 항체 용액 300 ul를 반응 용기에 넣고 상온에서 30분 동안 교반하였다. 항체용액을 버리고 세정액을 0.4 ml 분주하고 상온에서 5분간 교반한 후 제거하였다. Streptavidin-Alkaline Phosphatase (Streptavidin-AP) (Promeg, USA) 용액 0.4ml을 넣고 30분간 교반하였다. Streptavidin-AP용액을 버리고 세정액 (50mM 트리스, 0.2M NaCl, 0.05% Tween20) 0.4ml을 분주하고 상온에서 5분간 교반한 후 제거하였으며, 이 과정을 1회 추가하였다. 0.2mg/ml의 bromochlorophenyl phosphate와 0.3mg/ml의 nitroblue tetrazolium이 포함된 발색용액 400uL를 반응 용기에 넣고 상온에서 교반시키면서 발색반응을 시켰다. 20분 후 용액을 제거하고 400ul의 증류수를 넣고 세척하고 용액을 제거한 후 건조시켰다. 그 결과, 도 4에 나타난 바와 같이, 실험한 혈청 내의 특이 면역글로불린E(specific immunoglobulin E, s-IgE)가 있는 한약재 알레르겐에 대하여 검정색으로 발색됨을 확인할 수 있었다. 도 4의 왼쪽 (-) 스트립은 한약재 알레르기 음성 환자의 혈청, 오른쪽의 (+) 스트립은 한약재 알레르기 양성 환자의 혈청으로 실험이 실시되었다. 대조군 S1, S2, S3, S4 마커와 표1과 동일한 목록의 44종 한약재 마커를 비교하였을 때에 양성 환자 혈청으로 실험이 실시된 바이오 칩에서 다수의 한약재 마커에 대해 발색 반응이 나타났으며, 이 때 대조군 과의 발색 정도의 비교를 통해 검체 내 한약재와 반응하는 특이 면역글로불린(항체, antibody)의 농도를 정량화 할 수 있다. 300 ul of a Biotin-labeled mouse anti-phosphorus IgE antibody solution was placed in a reaction vessel and stirred at room temperature for 30 minutes. The antibody solution was discarded and 0.4 ml of the washing solution was dispensed, stirred at room temperature for 5 minutes, and then removed. 0.4 ml of Streptavidin-Alkaline Phosphatase (Streptavidin-AP) (Promeg, USA) solution was added and stirred for 30 minutes. The Streptavidin-AP solution was discarded and 0.4 ml of a washing solution (50 mM Tris, 0.2 M NaCl, 0.05% Tween20) was dispensed, stirred at room temperature for 5 minutes, and then removed. 400uL of the coloring solution containing 0.2mg / ml bromochlorophenyl phosphate and 0.3mg / ml nitroblue tetrazolium was added to the reaction vessel, and the coloring reaction was carried out while stirring at room temperature. After 20 minutes, the solution was removed, 400ul of distilled water was added, washed, and the solution was removed and dried. As a result, as shown in Figure 4, it was confirmed that the black color for the herbal medicinal allergen with the specific immunoglobulin E (specific immunoglobulin E, s-IgE) in the tested serum. The left (-) strip of Figure 4 is the serum of the herbal medicine allergy negative patients, the (+) strip of the right was tested with the serum of the herbal medicine allergy positive patients. Comparing the control markers S1, S2, S3, and S4 with the 44 kinds of herbal markers in the same list as in Table 1, color reactions were observed for a number of herbal markers in biochips tested with positive patient sera. By comparing the color development with the control group, it is possible to quantify the concentration of specific immunoglobulin (antibody, antibody) that reacts with the herbal medicine in the sample.
상기 결과를 통하여, 본 발명의 한약재 알레르겐 코팅 멤브레인을 통해 생물학적 시료에 존재하는 특이 면역글로불린E(specific Immunoglobulin E, s-IgE)의 농도 측정이 가능함을 확인하였다.Through the above results, it was confirmed that it is possible to measure the concentration of specific immunoglobulin E (specific Immunoglobulin E, s-IgE) present in the biological sample through the herbal allergen coating membrane of the present invention.
실시예Example 3. 혈청 내  3. In serum IgGIgG 농도 측정 Concentration measurement
상기 실시예 1과 실시예 2를 따르되 실시예 2의 Biotin 표지 마우스 항-인 IgE 항체 용액 300 ul 대신 Biotin 표지 마우스 항-인 IgG 항체 용액 300 ul를 사용하여 30분 동안 교반하였다. 상기 과정 이외의 나머지 과정을 실시예 1, 실시예 2와 동일하게 실시하였으며, 그 결과 특이 면역글로불린G(specific immunoglobulin G, s-IgG)와 반응하는 한약재 마커에서 검정색으로 발색 됨을 확인할 수 있다. 실시예 2와 마찬가지로 (-) 스트립은 음성 환자의 혈청 실험군, (+) 스트립은 양성 환자의 혈청 실험군이며, 대조군 S1, S2, S3, S4 마커와 표 1을 참고하여 특정 한약재 마커와 반응하는 특이 면역글로불린G(specific immunoglobulin G, s-IgG)의 확인 및 정량이 가능하다.Following Example 1 and Example 2, but using 300 ul of the Biotin-labeled mouse anti-phosphorus IgG antibody solution instead of 300 ul of the Biotin-labeled mouse anti-phosphorus IgE antibody solution of Example 2 was stirred for 30 minutes. The rest of the procedure was carried out in the same manner as in Example 1 and Example 2, and as a result, it was confirmed that the medicinal herb markers reacted with specific immunoglobulin G (s-IgG) were colored black. As in Example 2, the (-) strip is a sera test group of negative patients, and the (+) strip is a sera test group of positive patients. Identification and quantification of specific immunoglobulin G (s-IgG) is possible.
상기 결과를 통해 본 발명이 생물학적 시료 내의 특이 면역글로불린G(specific Immunoglobulin G, IgG)또한 검출 가능함을 확인하였다.The results confirm that the present invention can also detect specific immunoglobulin G (specific immunoglobulin G, IgG) in biological samples.
부호의 설명Explanation of the sign
10 : 라인형 스트립10: line strip
20 : 용기20 container
30 : 마커30: marker

Claims (10)

  1. 한약재와 반응하는 생채 시료 내 특이 면역글로불린(항체, antibody)의 농도를 측정하여 한약재로 야기되는 알레르기를 진단하는 한약재 알레르기 진단용 바이오칩.A biochip for diagnosing allergy caused by herbal medicine by diagnosing allergy caused by herbal medicine by measuring the concentration of specific immunoglobulin (antibody, antibody) in a raw vegetable sample reacting with herbal medicine
  2. 제1항에 있어서, 상기 바이오칩은 하나의 반응 용기 내에 스트립(strip); 및The method of claim 1, wherein the biochip comprises: a strip in one reaction vessel; And
    상기 스트립을 고정하는 용기; 를 포함하는 것을 특징으로 하는 한약재 알레르기 진단용 바이오칩.A container holding the strip; Herbal medicine allergy diagnostic biochip, comprising a.
  3. 제2항에 있어서, 상기 스트립은 라인형, 도트형, 병렬식 라인형으로 이루어진 군으로부터 선택되는 것을 특징으로 하는 한약재 알레르기 진단용 바이오칩.The method of claim 2, wherein the strip is a herb allergy diagnostic biochip, characterized in that selected from the group consisting of a line type, a dot type, parallel line type.
  4. 제2항에 있어서, 상기 스트립은 한약재 알레르겐(allergen)을 마커로 포함하는 것을 특징으로 하는 한약재 알레르기 진단용 바이오칩.[Claim 3] The biochip of claim 2, wherein the strip comprises a herb allergen as a marker.
  5. 제4항에 있어서, 상기 한약재 알레르겐(allergen)은 갈근, 감초, 건칠, 녹용, 백출, 일당귀, 지구자, 창출, 천궁, 행인, 황기, 지모, 인삼, 회향, 시호, 익모초, 생지황, 치자, 후박, 황백초, 복령, 지실, 작약, 황금, 봉독, 창이자, 구기자, 산수유, 보골지, 현삼, 목통, 도인, 연교, 속단, 마인, 저령, 고본, 백지, 산약, 석창포, 세신, 오미자, 택사, 맥문동으로 이루어진 군 중 1종 이상 포함하는 것을 특징으로 하는 한약재 알레르기 진단용 바이오칩. The method of claim 4, wherein the herbal allergen (allergen) is black root, licorice, dried, antler, baekryeok, daily donkey, earth, creation, cheonggung, passersby, Astragalus, gimo, ginseng, fennel, shiho, motherwort, raw sulfur, gardenia, Huak, Hwangbaekcho, Fuling, Jisil, Peony, Golden, Bee Venom, Changza, Wolfberry, Cornus, Bogolji, Hyunsam, Kowloon, Doin, Yeongyo, Sokdan, Mine, Spirit, Kobon, White Paper, Sanctuary, Seokchangpo, Sesin, Omija, A medicinal herb allergy diagnostic biochip, characterized in that it comprises one or more of the group consisting of Taeksa, Macmundong.
  6. 제2항에 있어서, 상기 스트립은 지지체 위에 멤브레인(membrane)을 코팅하는 것을 특징으로 하는 한약재 알레르기 진단용 바이오칩.The method of claim 2, wherein the strip is a herbal medicine allergy diagnostic biochip, characterized in that for coating a membrane (membrane) on the support.
  7. 제6항에 있어서, 상기 멤브레인은 니트로셀룰로스, 나일론, 폴리플루오린화비닐리덴(PVDF), 유리 및 플라스틱으로 이루어진 군으로부터 선택되는 것을 특징으로 하는 한약재 알레르기 진단용 바이오칩.The method of claim 6, wherein the membrane is a herbal medicine allergy diagnostic biochip, characterized in that selected from the group consisting of nitrocellulose, nylon, polyvinylidene fluoride (PVDF), glass and plastic.
  8. 제1항의 진단용 바이오칩 마커에 생물학적 시료를 접촉시켜, 시료 내에 존재하는 특이 면역글로불린(항체, antibody)의 농도를 측정하는 한약재 알레르기 진단 방법. A method of diagnosing medicinal herbs in which the biological sample is contacted with the diagnostic biochip marker of claim 1 to measure the concentration of specific immunoglobulin (antibody, antibody) present in the sample.
  9. 제8항에 있어서, 상기 면역글로불린은 면역글로불린 E(Immunoglobulin E, IgE) 또는 면역글로불린 G(Immunoglobulin G, IgG)인 것을 특징으로 하는 한약재 알레르기 진단 방법. The method of claim 8, wherein the immunoglobulin is immunoglobulin E (Immunoglobulin E, IgE) or immunoglobulin G (Immunoglobulin G, IgG).
  10. 제8항에 있어서, 상기 생물학적 시료는 조직, 세포, 전혈, 혈정, 혈장, 타액, 뇌척수액, 및 뇨로 이루어진 군으로부터 선택되는 것을 특징으로 하는 한약재 알레르기 진단 방법.The method of claim 8, wherein the biological sample is selected from the group consisting of tissue, cells, whole blood, blood serum, plasma, saliva, cerebrospinal fluid, and urine.
PCT/KR2016/008376 2016-07-18 2016-07-29 Biochip for diagnosing medicinal herb allergy WO2018016670A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR1020160091014A KR20180009443A (en) 2016-07-18 2016-07-18 Biochip for diagnosing medicinal herbs allergy
KR10-2016-0091014 2016-07-18

Publications (1)

Publication Number Publication Date
WO2018016670A1 true WO2018016670A1 (en) 2018-01-25

Family

ID=60993130

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2016/008376 WO2018016670A1 (en) 2016-07-18 2016-07-29 Biochip for diagnosing medicinal herb allergy

Country Status (2)

Country Link
KR (1) KR20180009443A (en)
WO (1) WO2018016670A1 (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
KR102634321B1 (en) * 2022-12-08 2024-02-08 주식회사 파나큐라 Composition for diagnosing deer antler allergy comprising albumin as an active ingredient
KR102634320B1 (en) * 2022-12-08 2024-02-08 주식회사 파나큐라 Composition for diagnosing bee venom allergy comprising phospholipase A2 as an active ingredient

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020187520A1 (en) * 1992-11-28 2002-12-12 Birgit Anna Helm Allergen/ inflammatory testing and diagnosis
US20100129831A1 (en) * 2007-06-07 2010-05-27 Sarah Brenner Methods for diagnosing hypersensitivity reactions
KR20140111192A (en) * 2013-03-08 2014-09-18 (주)프로테옴텍 Parallel line biochip for multiplex diagnosis

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020187520A1 (en) * 1992-11-28 2002-12-12 Birgit Anna Helm Allergen/ inflammatory testing and diagnosis
US20100129831A1 (en) * 2007-06-07 2010-05-27 Sarah Brenner Methods for diagnosing hypersensitivity reactions
KR20140111192A (en) * 2013-03-08 2014-09-18 (주)프로테옴텍 Parallel line biochip for multiplex diagnosis

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
2005 *
TREUDLER, R.: "Update on in vitro allergy diagnostics", JDDG: JOURNAL DER DEUTSCHEN DERMATOLOGISCHEN GESELLSCHAFT, 2012 *

Also Published As

Publication number Publication date
KR20180009443A (en) 2018-01-29

Similar Documents

Publication Publication Date Title
DE60108256T3 (en) ALLERGEN ASSAY BASED ON MICRO ASSEMBLIES
US6764849B2 (en) Rapid diagnostic method for distinguishing allergies and infections and nasal secretion collection unit
DE60120067T2 (en) Detection of multiple analytes in a single sample using a multi-vessel, multi-analyte, flow-through diagnostic test element
CN105408750B (en) Device and its application method is separated by electrophoresis
WO2014137069A1 (en) Multi-diagnosis parallel-type linear biochip
RU2397178C1 (en) Diagnostic test system in immunochip format and differential serum diagnostics of syphilis
WO2018016670A1 (en) Biochip for diagnosing medicinal herb allergy
KR101570327B1 (en) Parallel line biochip for multiplex diagnosis
CN109655608A (en) A kind of excretion body protein for osteosarcoma diagnosis and its instant detection method
EP2282206B1 (en) Method and apparatus for assaying several analytes simultaneously with an internal control
CN109239363A (en) A kind of application of the agglutinin probe combination in terms of identifying Activity budget gender based on Urine proteins sugar-type
EP2210097B1 (en) Method for the in vitro diagnosis and/or in-vitro therapy monitoring of infections
CN1300588C (en) Method for acquiring plasma specific protein for schizophrenia diagnosis and use thereof
DE60304336T2 (en) SERIOUS IN VITRO PROCEDURE FOR THE DIAGNOSIS OF INFECTIOUS ENDOCARDITIS
EP2035833B1 (en) Biochip and method for the selective identification of chlamydia trachomatis infections
Комар et al. SEROLOGICAL METHODS IN MODERN BIOTECHNOLOGIES AND THEIR BIOANALYTICAL STANDARDIZATION
EP3875957A1 (en) Method and device for diagnosing a suspected viral infection
Komar et al. Serological methods in modern biotechnologies and their bioanalytical standardization
EA024771B1 (en) Method for rapid laboratory diagnostics of diseases based on identification of specific proteins
Wolcott A study on the seroconversion rates in an area endemic for Histoplasma capsulatum
DE202005020150U1 (en) Reference material for quality control of antibody index measurements, useful in diagnostic determination of immunoglobulin levels against neurotropic pathogens, comprises serum and liquor cerebrospinalis pairs

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 16909585

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 16909585

Country of ref document: EP

Kind code of ref document: A1