An optical sensor for the detection of human pancreatic lipase

• Published in: Sensors and actuators. B, Chemical Chemical, 2014-11, Vol.203, p.663-669
• Main Authors: Krieg, A.K., Gauglitz, G.
• Format: Article
• Language: English
• Subjects: Fluorescence labeling; Human pancreatic lipase; Optical sensors; Reflectometric interference spectroscopy; Total internal reflection fluorescence
• ISSN: 0925-4005; 1873-3077
• DOI: 10.1016/j.snb.2014.07.036

•Development of a new heterogeneous immunoassay for human pancreas lipase.•Total internal reflection fluorescence was used as optical measuring method.•Good surface stability allows up to 50 measurements on each transducer.•Calibration range covers the clinically interesting concentration range. We present the development of a new heterogeneous immunoassay to determine the concentration of human pancreatic lipase (HPL) using reflectometric interference spectroscopy (RIfS) and total internal reflection fluorescence (TIRF). HPL is a digestion enzyme used as a biomarker to detect an acute inflammation of the pancreas. The assay is carried out using the binding inhibition assay format. RIfS is used for antibody characterization and assay development as with this method binding signals can be observed time-resolved. A good surface stability and reproducibility of the HPL biosensor can be realized. Using a suitable regeneration agent makes it possible to perform up to 50 measurements on each RIfS transducer. To achieve the required limit of detection, the immunoassay is transferred to the TIRF principle and is therefore the first HPL immunoassay based on fluorescence labeling of the antibody. A series of concentration measurements and their calibration show a sufficient working range of 0.068–3.849mg/L HPL using TIRF. On each TIRF transducer, 20–30 measurements can be performed.