Title:
METHOD FOR IDENTIFYING BHS-SPECIFIC PROTEINS AND FRAGMENTS THEREOF
Document Type and Number:
WIPO Patent Application WO2004029631
Kind Code:
A3
Abstract:
The invention relates to a method for identifying the presence of a BHS-specific protein/fragment in cerebral capillary endothelial cells, characterized in that a) cerebral endothelial cells freshly isolated from the brain are pre-cleaned by enzymatic decomposition in the usual manner; b) the decomposition obtained in step a) is treated with a lysis buffer which substantially destroys the erythrocytes and apoptic cells and enables at least 70 % of the cerebral capillary endothelial cells to be obtained in a vital form; c) optionally, the product obtained in step b) is further cleaned; d) a substractive cDNA bank is produced from the cerebral capillary endothelial cells and a substraction tissue; e) a cDNA substraction is performed by means of differential hybridization(s); f) clones from the substractive cDNA bank are verified with regard to the respective expression thereof by differential hybrization; g) a full cDNA sequence is established for the BHS-specific clones from the substractive cDNA bank and h) the expression pattern of the analysed clone is compared in relation to fresh and cultivated cerebral capillary endothelial cells, whereupon the presence of BHS-specific proteins/fragments are identified in addition to the proteins and fragments thereof identified according to said method.
Inventors:
WOLF SABINE (DE)
JAEGER MARTINA (DE)
BANGSOW THORSTEN (DE)
BANGSOW CARMEN (DE)
JORDAN DOMINIK (DE)
PELZER BERNHARD (DE)
OPPOLZER THOMAS (DE)
JAEGER MARTINA (DE)
BANGSOW THORSTEN (DE)
BANGSOW CARMEN (DE)
JORDAN DOMINIK (DE)
PELZER BERNHARD (DE)
OPPOLZER THOMAS (DE)
Application Number:
PCT/EP2003/009968
Publication Date:
March 02, 2006
Filing Date:
September 08, 2003
Export Citation:
Assignee:
FRANKGEN BIOTECHNOLOGIE AG (DE)
WOLF SABINE (DE)
JAEGER MARTINA (DE)
BANGSOW THORSTEN (DE)
BANGSOW CARMEN (DE)
JORDAN DOMINIK (DE)
PELZER BERNHARD (DE)
OPPOLZER THOMAS (DE)
WOLF SABINE (DE)
JAEGER MARTINA (DE)
BANGSOW THORSTEN (DE)
BANGSOW CARMEN (DE)
JORDAN DOMINIK (DE)
PELZER BERNHARD (DE)
OPPOLZER THOMAS (DE)
International Classes:
C07K14/47; A61K38/00; (IPC1-7): G01N33/68
Domestic Patent References:
| WO2001068848A2 | 2001-09-20 | |||
| WO2001073003A1 | 2001-10-04 | |||
| WO2003068054A2 | 2003-08-21 |
Other References:
DELEERSNIJDER W ET AL: "ISOLATION OF MARKERS FOR CHONDRO-OSTEOGENIC DIFFERENTIATION USING CDNA LIBRARY SUBTRACTION MOLECULAR CLONING AND CHARACTERIZATION OF A GENE BELONGING TO A NOVEL MULTIGENE FAMILY OF INTEGRAL MEMBRANE PROTEINS", JOURNAL OF BIOLOGICAL CHEMISTRY, AMERICAN SOCIETY OF BIOLOGICAL CHEMISTS, BALTIMORE, MD, US, vol. 271, no. 32, 9 August 1996 (1996-08-09), pages 19475 - 19482, XP002917436, ISSN: 0021-9258
CHOI S C ET AL: "Cloning and characterization of a type II integral transmembrane protein gene, Itm2c, that is highly expressed in the mouse brain.", MOLECULES AND CELLS. KOREA (SOUTH) 31 DEC 2001, vol. 12, no. 3, 31 December 2001 (2001-12-31), pages 391 - 397, XP009026835, ISSN: 1016-8478
SEN T ET AL: "Glucose oxidase-induced lysis of erythrocytes", INDIAN JOURNAL OF EXPERIMENTAL BIOLOGY, vol. 33, no. 1, 1995, pages 75 - 76, XP009026651, ISSN: 0019-5189
AUDUS K L ET AL: "Bovine brain microvessel endothelial cell monolayers as a model system for the blood-brain barrier.", ANNALS OF THE NEW YORK ACADEMY OF SCIENCES. UNITED STATES 1987, vol. 507, 1987, pages 9 - 18, XP001179819, ISSN: 0077-8923
YAO XIAO-HAO ET AL: "Identification of a key protein associated with cerebral ischemia.", BRAIN RESEARCH, vol. 967, no. 1-2, 28 March 2003 (2003-03-28), pages 11 - 18, XP001180022, ISSN: 0006-8993 (ISSN print)
CHOI S C ET AL: "Cloning and characterization of a type II integral transmembrane protein gene, Itm2c, that is highly expressed in the mouse brain.", MOLECULES AND CELLS. KOREA (SOUTH) 31 DEC 2001, vol. 12, no. 3, 31 December 2001 (2001-12-31), pages 391 - 397, XP009026835, ISSN: 1016-8478
SEN T ET AL: "Glucose oxidase-induced lysis of erythrocytes", INDIAN JOURNAL OF EXPERIMENTAL BIOLOGY, vol. 33, no. 1, 1995, pages 75 - 76, XP009026651, ISSN: 0019-5189
AUDUS K L ET AL: "Bovine brain microvessel endothelial cell monolayers as a model system for the blood-brain barrier.", ANNALS OF THE NEW YORK ACADEMY OF SCIENCES. UNITED STATES 1987, vol. 507, 1987, pages 9 - 18, XP001179819, ISSN: 0077-8923
YAO XIAO-HAO ET AL: "Identification of a key protein associated with cerebral ischemia.", BRAIN RESEARCH, vol. 967, no. 1-2, 28 March 2003 (2003-03-28), pages 11 - 18, XP001180022, ISSN: 0006-8993 (ISSN print)
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