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Development of a plasma panel test for detection of human myocardial proteins by capillary immunoassay
Uppsala universitet, Medicinska och farmaceutiska vetenskapsområdet, Medicinska fakulteten, Institutionen för kirurgiska vetenskaper, Rättsmedicin.
2007 (engelsk)Inngår i: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 22, nr 7, s. 1218-1223Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

A chemiluminescence immunoassay for the detection of four heart marker proteins: myoglobin, creatine kinase mb [CKmb], troponin I [TnI], and fatty acid-binding protein [FABP], was designed. The immunoassay was based on enzyme-linked immunosorbent assay [ELISA] and antibodies immobilized in glass capillaries pre-treated with 3-aminopropyltriethoxysilane. The protein bound to the antibody was detected by using an anti-protein-horseradish peroxidase [HRP] conjugate. The reaction of the HRP with luminal and hydrogen peroxide-based substrate generated the chemiluminescence and a photodiode detector was used to measure the light intensity. The same assay protocol was used to detect all four proteins. Ultrasound waves were used to improve the silanization of glass and the antibody immobilization process. The optimization of the duration and intensity of the ultrasound was performed for the myoglobin assay. Ultrasound improved the silanization procedure and the capillaries gave an approximately 2.5 times greater ELISA response. Ultrasound also improved the sensitivity by approximately 100% when monoclonal antibody was immobilized on a glass capillary. Calibration curves corresponding to analyte concentrations ranging from 2.4 to 2400 ng/ml in plasma samples were recorded. The detection limits were in the region of 1.2 myoglobin, 0.6 CKmb, 5.6 TnI, and 4 ng/ml FABP in plasma with a coefficient of variation of 3–9.9%.

sted, utgiver, år, opplag, sider
2007. Vol. 22, nr 7, s. 1218-1223
Emneord [en]
Chemiluminescent ELISA, Creatine kinase mb, Fatty acid-binding protein, Myoglobin, Troponin I, Ultrasound
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Identifikatorer
URN: urn:nbn:se:uu:diva-15375DOI: 10.1016/j.bios.2006.04.030ISI: 000244385400005PubMedID: 16797176OAI: oai:DiVA.org:uu-15375DiVA, id: diva2:43146
Tilgjengelig fra: 2008-02-11 Laget: 2008-02-11 Sist oppdatert: 2017-12-08bibliografisk kontrollert

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