title: α-Amylase sensor based on the degradation of oligosaccharide hydrogel films monitored with a quartz crystal sensor.
abstract: α-Amylase hydrolyses starch molecules to produce smaller oligosaccharides and sugars. Amylases are of great importance in biotechnology and find application in fermentation, detergents, food and the paper industry. The measurement of α-amylase activity in serum and urine has been used in the diagnosis of acute pancreatitis. Salivary amylase has also been shown to be a stress indicator. Sensor coatings suitable for the detection of α-amylase activity have been developed. Oligosaccharides such as glycogen and amylopectin were spin-coated onto gold coated quartz crystals with a base frequency of 10 MHz. The films were subsequently cross-linked with hexamethylene diisocyanate. Film degradation was monitored with a quartz crystal microbalance (QCM) and electrochemical impedance measurements. The films were shown to be stable in phosphate buffered saline (PBS). Addition of α-amylase to the solution resulted in the rapid degradation of the films. The maximum rate of degradation was found to be strongly dependent on the amylase activity in the range typically found in serum when diagnosing pancreatitis (0.08-8 U/ml). Sensor responses in serum were found to be very similar to those obtained in buffer indicating the absence of non-specific binding.
authors.affiliation: School of Engineering and Materials Science, Queen Mary University of London, Mile End Road, London E1 4NS, United Kingdom.
authors.forename: Martin John
authors.initials: MJ
authors.lastname: Gibbs
authors.affiliation: School of Engineering and Materials Science, Queen Mary University of London, Mile End Road, London E1 4NS, United Kingdom.
authors.forename: Anna
authors.initials: A
authors.lastname: Biela
authors.affiliation: School of Engineering and Materials Science, Queen Mary University of London, Mile End Road, London E1 4NS, United Kingdom. Electronic address: s.krause@qmul.ac.uk.
authors.forename: Steffi
authors.initials: S
authors.lastname: Krause
mesh_terms: D015374:Biosensing Techniques; D006801:Humans; D008756:Methylgalactosides / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry; D010944:Plants / Q000737:chemistry; D058525:Quartz Crystal Microbalance Techniques; D000516:alpha-Amylases / Q000737:chemistry / Q000302:isolation & purification*
chemical_list: D008756:Methylgalactosides; D009844:Oligosaccharides; C109801:hydrogel film; D000516:alpha-Amylases
keywords: Film degradation; Impedance; QCM; Sensor; α-Amylase activity