title: Development of a nanogold-based immunochromatographic assay for detection of morphine in urine using the Amor-HK16 monoclonal antibody.
abstract: A simple, rapid competitive immunochromatography (ICG) strip test was developed to detect morphine in urine samples using a monoclonal antibody produced in-house and conjugated to gold nanoparticles. Hybridoma cells were cultured and the Amor-HK16 monoclonal antibody against morphine was obtained from the supernatant after purification by salting out and passing through a Protein G-Agarose affinity column. Morphine was obtained from morphine sulfate and a C6-hemisuccinate derivative of morphine was prepared, conjugated to bovine serum albumin, and immobilized to a nitrocellulose membrane as the test line. Goat anti-mouse antibody was used as a binder in the control line in the detection zone of the strip. Colloidal gold particles of diameter approximately 20 nm were prepared and conjugated to the monoclonal antibody. The detection limit of the test strip was found to be 2000 ng/mL of morphine in urine samples. Reliability was determined by performing the ICG test on 103 urine samples and comparing the results with those obtained by thin-layer chromatography. The sensitivity of the test was 100%, and the analysis time for the assay was approximately 5 min. The new ICG method was adequately sensitive and accurate for the rapid screening of morphine in urine.
authors.affiliation: Department of Biochemistry, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Ardeshir
authors.initials: A
authors.lastname: Dehghannezhad
authors.forename: Maliheh
authors.initials: M
authors.lastname: Paknejad
authors.forename: Mohammad Javad
authors.initials: MJ
authors.lastname: Rasaee
authors.forename: Kobra
authors.initials: K
authors.lastname: Omidfar
authors.forename: Shadi Sadat
authors.initials: SS
authors.lastname: Seyyed Ebrahimi
authors.forename: Hossein
authors.initials: H
authors.lastname: Ghahremani
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D003101:Collodion / Q000737:chemistry; D017979:Gold Colloid / Q000737:chemistry*; D006801:Humans; D006825:Hybridomas; D006863:Hydrogen-Ion Concentration; D007159:Immunologic Tests; D057230:Limit of Detection; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D009020:Morphine / Q000276:immunology / Q000652:urine*; D010316:Particle Size; D012015:Reference Standards; D015203:Reproducibility of Results; D015813:Substance Abuse Detection / Q000379:methods* / Q000592:standards
chemical_list: D000911:Antibodies, Monoclonal; D017979:Gold Colloid; D009020:Morphine; D003101:Collodion