Human brain natriuretic peptide (hBNP) was used clinically for the treatment of acute decompensated congestive heart failure. In this paper, hBNP was expressed as a fusion protein with a histidine tag and Ssp dnaB mini-intein which was capable of self-cleavage. After affinity chromatography with Ni-Sepharose and renaturation, the fusion protein was enriched with CM-cellulose. Ssp dnaB mini-intein mediated peptide-bond hydrolysis was triggered by shifting the pH and temperature in the CM-cellulose column, which let to the release of hBNP from the fusion protein and the separation of hBNP from His-DnaB. The hBNP sample was further purified by C4 reverse phase HPLC, and 2.8mg of the peptide with homogeneity of 97% was obtained from one liter of culture medium. The biological activity was assayed in vitro, which indicated that hBNP had a potent vasodilatory effect on rabbit aortic strips with an EC50 of 1.94 x 10(-6) mg/mL.
Animals ; DnaB Helicases ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Inteins ; genetics ; Natriuretic Peptide, Brain ; biosynthesis ; genetics ; Protein Engineering ; methods ; Protein Splicing ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology

A kind of real-time remote monitoring embedded terminal which is combined with mobile communication technology and GPS localization technology, has been developed. The results of preliminary experiments show that the terminal can transmit ECG signals and localization information in real time and continuously, supply a real-time monitoring of out-of-hospital cardiac patients and trace the patients.
Electrocardiography, Ambulatory ; instrumentation ; Telemedicine ; instrumentation ; Telemetry ; instrumentation