کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1935871 1050676 2008 5 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Development of homogeneous immunoassays based on protein fragment complementation
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Development of homogeneous immunoassays based on protein fragment complementation
چکیده انگلیسی

We demonstrate a functional in vitro proof-of-principle homogeneous assay capable of detecting small (<1 kDa) to large (150 kDa) analytes using TEM-1 β-lactamase protein fragment complementation. In the assays reported here, complementary components are added together in the presence of analyte and substrate resulting in colorimetric detection within 10-min. We demonstrate the use of functional mutations leading to either increased enzymatic activity, reduced fragment self-association or increased inhibitor resistance upon analyte driven fragment complementation. Kinetic characterization of the resulting reconstituted enzyme illustrates the importance of balancing increased enzyme activity with fragment self-association, producing diagnostically relevant signal-to-noise ratios. Complementation can be utilized as a homogeneous immunoassay platform for the potential detection of a range of analytes including, antibodies, antigens and biomarkers.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Biochemical and Biophysical Research Communications - Volume 370, Issue 1, 23 May 2008, Pages 164–168
نویسندگان
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