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Development of a recombinant immunotoxin for the immunotherapy of autoreactive lymphocytes expressing MOG-specific BCRs

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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.

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Acknowledgments

This work was supported by the Russian Scientific Foundation grant 14-24-00106. We thank Dr. Yury Belyi (Gamaleya Research Institute, Russia) for a plasmid coding for the A-subunit of DT. A.S. obtained personal support from RFBR, research project No. 15-34-70037 mol_a_mos.

Supporting information

Supplementary Table 1—Optimization of the recombinant immunotoxin expression in pET-28 vector.

Supplementary Table 2—The immunotoxin purification summary table.

Supplementary Table 3—Acute in vivo toxicity of a single i.v. injection of DT-MOG in mice.

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Correspondence to Alexander Gabibov.

Additional information

Alexey Stepanov and Alexander Belyy contributed equally to this work.

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Stepanov, A., Belyy, A., Kasheverov, I. et al. Development of a recombinant immunotoxin for the immunotherapy of autoreactive lymphocytes expressing MOG-specific BCRs. Biotechnol Lett 38, 1173–1180 (2016). https://doi.org/10.1007/s10529-016-2092-5

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  • DOI: https://doi.org/10.1007/s10529-016-2092-5

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