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Antibodies 2013, 2(1), 93-112; doi:10.3390/antib2010093

Characterization of a Single Chain Fv Antibody that Reacts with Free Morphine

1 Department of Chemistry, Biotechnology and Chemical Engineering, Graduate School of Science and Engineering, Kagoshima University, 1-21-24 Korimoto, Kagoshima, Kagoshima 890-8580, Japan 2 Genenet CO., LTD. 5-22-8 Tanotsu, Higashi-ku, Fukuoka 813-0034, Japan 3 Faculty of Environmental Studies, Nagasaki University, 1-14 Bunkyou-machi, Nagasaki 852-8521, Japan 4 Faculty of Environmental and Symbiotic Sciences, Prefectural University of Kumamoto, 3-1-100 Tsukide, Kumamoto 862-8502, Japan
* Author to whom correspondence should be addressed.
Received: 10 December 2012 / Revised: 9 January 2013 / Accepted: 5 February 2013 / Published: 7 February 2013
(This article belongs to the Special Issue Single-Domain Antibody)
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An immune phage library derived from mice, hyperimmunized with morphine-conjugated BSA, was used to isolate a single-chain Fv (scFv) clone, M86, with binding activity to morphine-conjugated thyroglobulin (morphine-C-Tg) but not to codeine-, cocaine-, or ketamine-conjugated Tg. Surface plasmon resonance analysis using a morphine-C-Tg-coupled CM5 sensor chip showed that the Kd value was 1.26 × 108 M. To analyze its binding activity to free morphine and related compounds, we performed a competitive ELISA with M86 and morphine-C-Tg in the absence or presence of varying doses of free morphine and related compounds. IC50 values for opium, morphine, codeine, and heroin were 257 ng/mL, 36.4, 7.3, and 7.4 nM, respectively. Ketamine and cocaine exhibited no competitive binding activity to M86. Thus, we established a phage library-derived scFv, M86, which recognized not only free morphine and codeine as opium components but also heroin. This characteristic of M86 may be useful for developing therapeutic reagents for opiate addiction and as a free morphine-specific antibody probe.
Keywords: morphine; narcotic; hapten; single-chain Fv (scFv); recombinant antibody technology morphine; narcotic; hapten; single-chain Fv (scFv); recombinant antibody technology
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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Matsukizono, M.; Kamegawa, M.; Tanaka, K.; Kohra, S.; Arizono, K.; Hamazoe, Y.; Sugimura, K. Characterization of a Single Chain Fv Antibody that Reacts with Free Morphine. Antibodies 2013, 2, 93-112.

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