Stepanov Alexey, Belyy Alexander, Kasheverov Igor, Rybinets Alexandra, Dronina Maria, Dyachenko Igor, Murashev Arkady, Knorre Vera, Sakharov Dmitry, Ponomarenko Natalya, Tsetlin Victor, Tonevitsky Alexander, Deyev Sergey, Belogurov Alexey, Gabibov Alexander
M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Str., 16/10, Moscow, Russia, 117997.
Institute of Fundamental Medicine and Biology, Kazan Federal University, Kazan, Tatarstan, Russia.
Biotechnol Lett. 2016 Jul;38(7):1173-80. doi: 10.1007/s10529-016-2092-5. Epub 2016 Apr 21.
Myelin oligodendrocyte glycoprotein (MOG) is one of the major autoantigens in multiple sclerosis (MS), therefore selective depletion of autoreactive lymphocytes exposing MOG-specific B cell receptors (BCRs) would be beneficial in terms of MS treatment.
Using E. coli we generated an efficient protocol for the purification of the recombinant immunotoxin DT-MOG composed of the extracellular Ig-like domain of MOG fused in frame with the catalytic and translocation subunits of diphtheria toxin (DT, Corynebacterium diphtheriae) under native conditions with a final yield of 1.5 mg per liter of culture medium. Recombinant DT-MOG was recognized in vitro by MOG-reactive antibodies and has catalytic activity comparable with wild-type DT.
Enhanced pharmacokinetics (mean residence time in the bloodstream of 61 min) and minimized diminished nonspecific toxicity (LD50 = 1.76 mg/kg) of the DT-MOG makes it a potential candidate for the immunotherapy of MS.
髓鞘少突胶质细胞糖蛋白（MOG）是多发性硬化症（MS）的主要自身抗原之一，因此，选择性清除暴露MOG特异性B细胞受体（BCR）的自身反应性淋巴细胞对MS治疗有益。
我们利用大肠杆菌建立了一种高效的重组免疫毒素DT-MOG纯化方案，该免疫毒素由MOG的细胞外Ig样结构域与白喉毒素（DT，白喉棒状杆菌）的催化亚基和转位亚基读码框融合而成，在天然条件下每升培养基最终产量为1.5毫克。重组DT-MOG在体外被MOG反应性抗体识别，并且具有与野生型DT相当的催化活性。
DT-MOG增强的药代动力学（在血液中的平均驻留时间为61分钟）和最小化的非特异性毒性（半数致死量=1.76毫克/千克）使其成为MS免疫治疗的潜在候选药物。
