RU2004114213A - Функциональный анализ активации рецепторов агонистами - Google Patents
Функциональный анализ активации рецепторов агонистами Download PDFInfo
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- RU2004114213A RU2004114213A RU2004114213/15A RU2004114213A RU2004114213A RU 2004114213 A RU2004114213 A RU 2004114213A RU 2004114213/15 A RU2004114213/15 A RU 2004114213/15A RU 2004114213 A RU2004114213 A RU 2004114213A RU 2004114213 A RU2004114213 A RU 2004114213A
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- G—PHYSICS
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- 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
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- 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
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
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- 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
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
-
- 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
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
- G01N33/5058—Neurological cells
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- 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
- G01N33/6872—Intracellular protein regulatory factors and their receptors, e.g. including ion channels
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- 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
- G01N33/94—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors
- G01N33/9406—Neurotransmitters
- G01N33/9413—Dopamine
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- 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
- G01N33/94—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors
- G01N33/9406—Neurotransmitters
- G01N33/942—Serotonin, i.e. 5-hydroxy-tryptamine
-
- 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
- G01N33/94—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving narcotics or drugs or pharmaceuticals, neurotransmitters or associated receptors
- G01N33/9406—Neurotransmitters
- G01N33/9433—(Nor)adrenaline
-
- 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
- G01N2333/72—Assays involving receptors, cell surface antigens or cell surface determinants for hormones
- G01N2333/726—G protein coupled receptor, e.g. TSHR-thyrotropin-receptor, LH/hCG receptor, FSH
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Cell Biology (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Tropical Medicine & Parasitology (AREA)
- Toxicology (AREA)
- Pharmacology & Pharmacy (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Heterocyclic Carbon Compounds Containing A Hetero Ring Having Oxygen Or Sulfur (AREA)
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
Claims (11)
1. Способ анализа для определения активации агонистом рецептора, связанного с G-белком, причем указанный способ основан на применении флуориметрического сцинтилляционного планшет-ридера, который предусматривает
(а) получение клеточной линии, характеризующейся, по крайней мере, одним подходящим фактором селекции, выбранным из маркера резистентности к лекарству, выбранного из НЕК293-G альфа 15, причем указанная клеточная линия стабильно экспрессирует смешанный G-белок, выбранный из G альфа 15, и, кроме того, коэкспрессирует в указанной клеточной линии упомянутый G-связанный рецептор, в результате трансфекции названной клеточной линии кДНК, кодирующей выбранный G-связанный рецептор;
(b) выращивание коэкспрессирующих клеток в соответствующей среде;
(с) содержание указанных клеток в чашках приблизительно в течение одного дня;
(d) нанесение на содержащиеся в чашках клетки некоторого количества флуоресцентной краски, соответствующей данной цели;
(е) инкубацию покрытых краской клеток при температуре приблизительно от комнатной температуры до 37°С приблизительно в течение подходящего периода;
(f) промывание планшета для удаления избытка краски подходящим буфером и замену объема удаленного буфера таким же объемом свежего буфера;
(g) инкубацию при температуре приблизительно от 30 до 37°С;
(h) добавление агониста при постоянных температурных условиях приблизительно от 30 до 37°С; и
(i) измерение испускания флуоресценции при постоянных температурных условиях приблизительно от 30 до 37°С во флуориметрическом сцинтилляционном планшет-ридере с тем, чтобы определить уровень активации выбранного рецептора агонистическим соединением.
2. Способ по п.1, при котором указанный G-связанный рецептор является дофаминовым или гистаминовым рецептором.
3. Способ по п.1, при котором указанный G-связанный рецептор выбирают из группы, состоящей из рецепторов D2, D3, альфа 1А, альфа 2А, М1, Н1, 5НТ1А и 5НТ2А.
4. Способ по п.1, при котором указанный G-связанный рецептор представляет собой дофаминовый рецептор D3.
5. Способ по п.1, при котором указанный фактор селекции, выбранный из маркера резистентности к лекарству, представляет собой маркер резистентности к пуромицину.
6. Способ по п.1, при котором указанный фактор селекции, выбранный из маркера резистентности к лекарству, представляет собой маркер резистентности к бластоцидину.
7. Способ по п.1, при котором указанный флуоресцентный краситель представляет собой Fluo-3TM или Fluo-4TM.
8. Способ по п.1, при котором высеянные в чашки клетки имеют плотность приблизительно между 12000 и 30000 клеток/квадратный см.
9. Способ по п.1, при котором указанную стадию инкубации (е) осуществляют приблизительно в течение одного часа.
10. Способ по п.1, при котором указанную стадию инкубации (g) осуществляют приблизительно в течение от 15 до 60 мин.
11. Способ по п.1, при котором указанную стадию инкубации (g) осуществляют приблизительно в течение 30 мин.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US34475501P | 2001-11-09 | 2001-11-09 | |
| US60/344,755 | 2001-11-09 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| RU2004114213A true RU2004114213A (ru) | 2005-03-20 |
Family
ID=23351886
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| RU2004114213/15A RU2004114213A (ru) | 2001-11-09 | 2002-10-30 | Функциональный анализ активации рецепторов агонистами |
Country Status (12)
| Country | Link |
|---|---|
| US (1) | US20030104489A1 (ru) |
| EP (1) | EP1310800A3 (ru) |
| JP (1) | JP3676774B2 (ru) |
| KR (1) | KR20040062623A (ru) |
| CN (1) | CN1774630A (ru) |
| AU (1) | AU2002350999A1 (ru) |
| CA (1) | CA2411476A1 (ru) |
| HR (1) | HRP20040407A2 (ru) |
| IL (1) | IL161695A0 (ru) |
| NO (1) | NO20041883L (ru) |
| RU (1) | RU2004114213A (ru) |
| WO (1) | WO2003040303A2 (ru) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB0421693D0 (en) | 2004-09-30 | 2004-11-03 | Amersham Biosciences Uk Ltd | Method for measuring binding of a test compound to a G-protein coupled receptor |
| CN101063657A (zh) * | 2006-04-29 | 2007-10-31 | 中国科学院上海生命科学研究院 | 在活体细胞中筛选配体与受体结合的方法和系统 |
| KR100852284B1 (ko) * | 2006-07-07 | 2008-08-14 | 한국과학기술연구원 | 형광 이미징법을 이용한 ５－ｈｔ６ 수용체 리간드 고효율검색법 |
| DE102007011913A1 (de) * | 2007-03-13 | 2008-10-23 | Sanofi-Aventis | Fluoreszenz-basiertes Assay zum Erkennen von Verbindungen zum Modulieren des Natrium-Calcium-Austauschers (NCX) im "Vorwärtsmodus" |
| US8551718B2 (en) | 2009-12-29 | 2013-10-08 | Suven Life Science Limited | Functional assay for 5-HT2A, histamine H1 or adrenergic alpha 1B receptors |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1096009B1 (en) * | 1999-10-29 | 2007-10-10 | Pfizer Limited | G-protein coupled receptor-like Polypeptide |
-
2002
- 2002-10-30 IL IL16169502A patent/IL161695A0/xx unknown
- 2002-10-30 CN CNA028268504A patent/CN1774630A/zh active Pending
- 2002-10-30 RU RU2004114213/15A patent/RU2004114213A/ru not_active Application Discontinuation
- 2002-10-30 KR KR10-2004-7006963A patent/KR20040062623A/ko not_active Application Discontinuation
- 2002-10-30 AU AU2002350999A patent/AU2002350999A1/en not_active Abandoned
- 2002-10-30 WO PCT/IB2002/004557 patent/WO2003040303A2/en not_active Application Discontinuation
- 2002-11-06 EP EP02257707A patent/EP1310800A3/en not_active Withdrawn
- 2002-11-07 US US10/289,818 patent/US20030104489A1/en not_active Abandoned
- 2002-11-07 CA CA002411476A patent/CA2411476A1/en not_active Abandoned
- 2002-11-08 JP JP2002325137A patent/JP3676774B2/ja not_active Expired - Fee Related
-
2004
- 2004-05-07 NO NO20041883A patent/NO20041883L/no not_active Application Discontinuation
- 2004-05-07 HR HR20040407A patent/HRP20040407A2/xx not_active Application Discontinuation
Also Published As
| Publication number | Publication date |
|---|---|
| EP1310800A3 (en) | 2004-02-11 |
| JP2003194810A (ja) | 2003-07-09 |
| AU2002350999A1 (en) | 2003-05-19 |
| US20030104489A1 (en) | 2003-06-05 |
| WO2003040303A3 (en) | 2003-10-09 |
| HRP20040407A2 (en) | 2005-06-30 |
| EP1310800A2 (en) | 2003-05-14 |
| NO20041883L (no) | 2004-06-01 |
| CN1774630A (zh) | 2006-05-17 |
| IL161695A0 (en) | 2004-09-27 |
| CA2411476A1 (en) | 2003-05-09 |
| KR20040062623A (ko) | 2004-07-07 |
| WO2003040303A2 (en) | 2003-05-15 |
| JP3676774B2 (ja) | 2005-07-27 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FA92 | Acknowledgement of application withdrawn (lack of supplementary materials submitted) |
Effective date: 20060113 |