A supercharged fluorescent protein based FRET sensing platform for detection of heparin contamination. Issue 38 (22nd September 2017)

• Record Type: Journal Article
• Authors: Ding, Yubin; Zhou, Min; Wei, Hui
• Is Part Of: Analytical methods. Volume 9:Issue 38(2017)
• Page Start: 5593
• Page End: 5597
• Publication Date: 2017-09-22
• Subjects: Chemistry, Analytic -- Periodicals; Analytical biochemistry -- Periodicals; Chemical laboratories -- Standards -- Periodicals; 543.1905
• DOI: 10.1039/c7ay01907f
• Languages: English
• ISSNs: 1759-9660

Abstract

Abstract : A FRET system was successfully constructed using supercharged green fluorescent protein as the energy donor and dye labelled heparin as the energy acceptor. The FRET sensing platform can be used to detect OSCS contaminant in heparin. Abstract : Development of biosensors for the detection of oversulfated chondroitin sulfate (OSCS), the key contaminant in heparin (Hep), is a greatly demanding but challenging task. In this work, a FRET system for the detection of OSCS was successfully constructed by using supercharged green fluorescent protein (ScGFP ) as the energy donor and dye labelled Hep (Hep-RF1 ) as the energy acceptor. With heparinase treatment, Hep-RF1 was hydrolyzed into small fragments, resulting in quenching of the FRET signal. On the other hand, since OSCS is an inhibitor for heparinase, the presence of OSCS would enable the effective FRET fromScGFP toHep-RF1 even after heparinase treatment. With thisScGFP based FRET sensing platform, as low as 0.001% (w/w) OSCS in Hep has been successfully detected.