کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1175652 961811 2009 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Development of an immobilized GPR17 receptor stationary phase for binding determination using frontal affinity chromatography coupled to mass spectrometry
کلمات کلیدی
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Development of an immobilized GPR17 receptor stationary phase for binding determination using frontal affinity chromatography coupled to mass spectrometry
چکیده انگلیسی

A liquid chromatographic stationary phase containing immobilized membranes from cells expressing the P2Y-like receptor GPR17 is described. Cellular membranes from 1321N1 cells transiently transfected with GPR17 vector [GPR17(+)] and from the same cell line transfected with the corresponding empty vector [GPR17(−)] were entrapped on immobilized artificial membrane (IAM) support and packed into 6.6-mm-i.d. glass columns to create GPR17(+)–IAM and GPR17(−)–IAM stationary phases. Frontal chromatography experiments on both GPR17(+)–IAM and GPR17(−)–IAM demonstrated the presence of a specific interaction with GPR17 only in the former that was maximized by increasing the membrane/IAM ratio. GPR17(+)–IAM was used in frontal affinity chromatography experiments to calculate the dissociation constants (Kd) of three ligands—the antagonist cangrelor (formerly AR-C69931MX, a P2Y12/P2Y13 antagonist), MRS2179 (a P2Y1 receptor antagonist), and the agonist UDP—all of which have been reported to also interact with GPR17. Immobilized GPR17 retained its ability to specifically bind the three analytes, as demonstrated by the agreement of the calculated Kd values with previously reported data. Preliminary ranking experiments suggest the application of GPR17(+)–IAM in ranking affinity studies for the selection of new potential candidates.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 384, Issue 1, 1 January 2009, Pages 123–129
نویسندگان
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