Oboznaia M B, Vladimirova N M, Titova M A, Volkova T D, Koroev D O, Riabokon' A A, Egorov A A, Rybakov S S, Vol'pina O M
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, ul. Miklukho-Maklaya 16/10, Moscow, 117997 Russia.
Bioorg Khim. 2008 Nov-Dec;34(6):754-63. doi: 10.1134/s1068162008060058.
Antibodies to the prion protein (PrP), particularly, monoclonal antibodies, are necessary tools in the diagnostics and study of prion diseases and potential means of their immunotherapy. For the production of monoclonal antibodies, BALB/c mice were immunized by a recombinant bovine PrP. Three stable hybridomas producing antibodies of IgM class were prepared. The antibodies were bound to PrP in a solid-phase enzyme immunoassay and immunoblotting. The epitope mapping accomplished with the use of synthetic peptides showed that an epitope located in region 25-36 of PrP corresponds to one antibody, and epitopes located in region 222-229, to the other two. The antibodies to fragment 222-229 purified by affinity chromatography recognized with a high specificity conglomerates of a pathogenic prion in the brain tissue of cows suffering from spongiform encephalopathy. Thus, in nontransgenic mice, PrP-specific monoclonal antibodies were produced, useful in studies and diagnostics of prion diseases.
抗朊病毒蛋白(PrP)抗体,尤其是单克隆抗体,是朊病毒疾病诊断和研究的必要工具,也是其免疫治疗的潜在手段。为了生产单克隆抗体,用重组牛PrP免疫BALB/c小鼠。制备了三种产生IgM类抗体的稳定杂交瘤。这些抗体在固相酶免疫测定和免疫印迹中与PrP结合。使用合成肽完成的表位作图表明,位于PrP 25-36区域的一个表位对应一种抗体,位于222-229区域的表位对应另外两种抗体。通过亲和层析纯化的针对222-229片段的抗体以高特异性识别患有海绵状脑病的奶牛脑组织中致病性朊病毒的聚集体。因此,在非转基因小鼠中产生了PrP特异性单克隆抗体,可用于朊病毒疾病的研究和诊断。