Alexey Stepanov1,2, Alexander Belyy1,3, Igor Kasheverov1, Alexandra Rybinets1, Maria Dronina1, Igor Dyachenko1, Arkady Murashev1, Vera Knorre1, Dmitry Sakharov4, Natalya Ponomarenko1, Victor Tsetlin1, Alexander Tonevitsky4, Sergey Deyev1, Alexey Belogurov1,2, Alexander Gabibov5,6. 1. M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Str., 16/10, Moscow, Russia, 117997. 2. Institute of Fundamental Medicine and Biology, Kazan Federal University, Kazan, Tatarstan, Russia. 3. Department of Bacterial Infections, Gamaleya Research Institute, Gamaleya Str., 18, Moscow, Russia, 123098. 4. SRC Bioclinicum, Ugreshskaya str 2/85, Moscow, Russia, 115088. 5. M.M. Shemyakin and Yu.A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya Str., 16/10, Moscow, Russia, 117997. gabibov@mx.ibch.ru. 6. Institute of Fundamental Medicine and Biology, Kazan Federal University, Kazan, Tatarstan, Russia. gabibov@mx.ibch.ru.
Abstract
OBJECTIVE: 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. RESULTS: 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. CONCLUSION: 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.
OBJECTIVE: 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. RESULTS: 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. CONCLUSION: 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.
Authors: Diana Klose; Mira Woitok; Judith Niesen; Roger R Beerli; Ulf Grawunder; Rainer Fischer; Stefan Barth; Rolf Fendel; Thomas Nachreiner Journal: PLoS One Date: 2017-07-13 Impact factor: 3.240