Moghaddam Amir, Borgen Tine, Stacy John, Kausmally Louise, Simonsen Bjørg, Marvik Ole J, Brekke Ole Henrik, Braunagel Michael
GeNova AS, Oslo Research Park, Guastadalléen 21, N-0349 Oslo, Norway.
J Immunol Methods. 2003 Sep;280(1-2):139-55. doi: 10.1016/s0022-1759(03)00109-1.
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.
利用重组抗体的噬菌体展示和大容量天然抗体文库来鉴定高特异性抗体的方法已被广泛报道。然而，源于天然抗体文库、具有诊断潜在用途的针对半抗原的抗体报道却很少。我们利用从天然抗体文库中分离出的先导单链可变片段（scFv）抗体进行链改组，以筛选特异性识别海洛因主要代谢物6-单乙酰吗啡（6MAM）的抗体。通过筛选表达可溶性scFv抗体片段的大肠杆菌高密度菌落来鉴定这些抗体，而无需事先在噬菌体上表达（噬菌体展示）。这些抗体识别6MAM的亲和力为1 - 3×10⁻⁷ M，对吗啡无交叉反应。这些抗体有可能用于开发药物检测项目中的快速免疫测定法。据我们所知，这是第一份关于区分6MAM与其脱乙酰化形式吗啡的抗体的报告。
