METHOD FOR IDENTIFYING BHS-SPECIFIC PROTEINS AND FRAGMENTS THEREOF

摘要

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.