Development of a plasma panel test for detection of human myocardial proteins by capillary immunoassay

Fereidon Torabi, Hamid Reza Mobini Far, Bengt Danielsson, Masoud Khayyami

Research output: Contribution to journalArticlepeer-review

Abstract

A chemiluminescence inummoassay for the detection of four heart marker proteins: myoglobin, creatine kinase mb [CKmb], troponin I [Tnl], and fatty acid-binding protein [FABP], was designed. The inummoassay 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 antiprotein-horseradish peroxidase [HR-P] 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%. (c) 2006 Elsevier B.V. All rights reserved.
Original languageEnglish
Pages (from-to)1218-1223
JournalBiosensors & Bioelectronics
Volume22
Issue number7
DOIs
Publication statusPublished - 2007

Subject classification (UKÄ)

  • Biochemistry and Molecular Biology

Free keywords

  • ultrasound
  • chemiluminescent ELISA
  • creatine kinase mb
  • myoglobin
  • fatty acid-binding protein
  • troponin I

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