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

机译：与游离吗啡反应的单链Fv抗体的表征

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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 × 10−8 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.
机译：使用小鼠吗啡结合BSA进行超免疫的免疫噬菌体文库用于分离单链Fv（scFv）克隆M86，其对吗啡结合甲状腺球蛋白（吗啡-C-Tg）具有结合活性，但对可待因没有结合活性-，可卡因或氯胺酮结合的Tg。使用吗啡-C-Tg偶联的CM5传感器芯片进行的表面等离子体共振分析表明K d 值为1.26×10 -8 M。游离吗啡及相关化合物，我们在不存在或存在不同剂量的游离吗啡及相关化合物的情况下，用M86和吗啡-C-Tg进行了竞争性ELISA。鸦片，吗啡，可待因和海洛因的IC 50 值分别为257 ng / mL，36.4、7.3和7.4 nM。氯胺酮和可卡因对M86没有竞争性结合活性。因此，我们建立了噬菌体文库衍生的scFv M86，它不仅将游离吗啡和可待因识别为鸦片成分，而且还识别了海洛因。 M86的这一特性可用于开发鸦片成瘾的治疗试剂，并用作游离的吗啡特异性抗体探针。

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