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Erythrocytes as bioreactors to decrease excess ammonium concentration in blood.

Eugeniy S Protasov1,2,3, Daria V Borsakova1,3, Yuliya G Alexandrovich1

  • 1Dmitriy Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology, Ministry of Healthcare, Samory Mashela str., 1, GSP-7, Moscow, 117997, Russia.

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|February 8, 2019
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Summary
This summary is machine-generated.

Researchers developed improved erythrocyte-bioreactors (EBRs) for removing toxic ammonium from blood. Mathematical models identified cell membrane permeability as a key limitation, leading to a novel enzyme system that enhances EBR efficiency and duration for treating hyperammoniemia.

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Area of Science:

  • Biomedical Engineering
  • Biotechnology
  • Metabolic Engineering

Background:

  • Elevated blood ammonium levels cause neurological issues, with current treatments often insufficient.
  • Erythrocyte-bioreactors (EBRs) using enzymes to clear ammonium were proposed but had limited functionality.
  • The reasons for short-term EBR function were previously uninvestigated.

Purpose of the Study:

  • To investigate the limitations of erythrocyte-bioreactors (EBRs) for ammonium removal.
  • To develop and validate mathematical models for EBRs.
  • To design and experimentally test a novel enzyme system to overcome EBR limitations.

Main Methods:

  • Developed and analyzed mathematical models of EBRs incorporating glycolysis and ammonium-utilizing enzymes.
  • Identified cell membrane permeability as a critical factor affecting substrate and product transport.
  • Designed and implemented a new enzyme system (glutamate dehydrogenase and alanine aminotransferase) with a cyclic metabolic pathway.

Main Results:

  • Mathematical models indicated that cell membrane permeability limits EBR efficiency and duration.
  • The novel enzyme system demonstrated efficient ammonium removal in vitro (1.5 mmol/h×lRBCs) and in vivo in a mouse hyperammoniemia model (2.0 mmol/h×lRBCs).
  • Experimental results validated the accuracy of the developed mathematical models.

Conclusions:

  • Cell membrane permeability is a significant limitation for conventional EBRs.
  • The proposed cyclic enzyme system effectively overcomes these permeability issues, enhancing ammonium removal.
  • Mathematical simulation of EBRs provides a powerful tool for analyzing and optimizing enzyme-based bioreactors.