title: Identification of scFv antibody fragments that specifically recognise the heroin metabolite 6-monoacetylmorphine but not morphine.
abstract: Use of phage display of recombinant antibodies and large repertoire naïve antibody libraries for identifying antibodies of high specificity has been extensively reported. Nevertheless, there have been few reported antibodies to haptens that have originated from naïve antibody libraries with potential use in diagnostics. We have used chain shuffling of lead single-chain fragment variable (scFv) antibodies, isolated from a naïve antibody library, to screen for antibodies that specifically recognise the major metabolite of heroin, 6-monoacetylmorphine (6MAM). The antibodies were identified by screening high-density colonies of Escherichia coli expressing soluble scFv antibody fragments without prior expression on bacteriophage (phage display). The antibodies recognise 6MAM with affinities of 1-3x10(-7) M with no crossreactivity to morphine. These antibodies can potentially be used for developing a rapid immunoassay in drug-testing programs. To our knowledge, this is the first report of an antibody that distinguishes 6MAM from its de-acetylated form, morphine.
authors.affiliation: GeNova AS, Oslo Research Park, Guastadalléen 21, N-0349 Oslo, Norway.
authors.forename: Amir
authors.initials: A
authors.lastname: Moghaddam
authors.forename: Tine
authors.initials: T
authors.lastname: Borgen
authors.forename: John
authors.initials: J
authors.lastname: Stacy
authors.forename: Louise
authors.initials: L
authors.lastname: Kausmally
authors.forename: Bjørg
authors.initials: B
authors.lastname: Simonsen
authors.forename: Ole J
authors.initials: OJ
authors.lastname: Marvik
authors.forename: Ole Henrik
authors.initials: OH
authors.lastname: Brekke
authors.forename: Michael
authors.initials: M
authors.lastname: Braunagel
mesh_terms: D000595:Amino Acid Sequence; D000918:Antibody Specificity; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D003932:Heroin / Q000378:metabolism*; D006801:Humans; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D009020:Morphine / Q000276:immunology; D009022:Morphine Derivatives / Q000276:immunology* / Q000378:metabolism*; D019151:Peptide Library; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D020349:Surface Plasmon Resonance
chemical_list: D007135:Immunoglobulin Variable Region; D009022:Morphine Derivatives; D019151:Peptide Library; D011994:Recombinant Proteins; D003932:Heroin; D009020:Morphine; C026979:6-O-monoacetylmorphine