Identification of scFv antibody fragments that specifically recognise the heroin metabolite 6-monoacetylmorphine but not morphine.
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.
Moghaddam
Amir
A
GeNova AS, Oslo Research Park, Guastadalléen 21, N-0349 Oslo, Norway.
Borgen
Tine
T
Stacy
John
J
Kausmally
Louise
L
Simonsen
Bjørg
B
Marvik
Ole J
OJ
Brekke
Ole Henrik
OH
Braunagel
Michael
M
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
D007135:Immunoglobulin Variable Region; D009022:Morphine Derivatives; D019151:Peptide Library; D011994:Recombinant Proteins; D003932:Heroin; D009020:Morphine; C026979:6-O-monoacetylmorphine