Expression and Purification of Neurotrophin-Elastin-Like Peptide Fusion Proteins for Neural Regeneration

BioDrugs : Clinical Immunotherapeutics, Biopharmaceuticals and Gene Therapy
Tamina Johnson, Piyush Koria

Abstract

Neural injuries such as spinal cord injuries, traumatic brain injuries, or nerve transection injuries pose a major health problem. Neurotrophins such as nerve growth factor (NGF) or brain-derived neurotrophic factor (BDNF) have been shown to improve the outcome of neural injuries in several pre-clinical models, but their use in clinics is limited by the lack of a robust delivery system that enhances their bioavailability and half-life. We describe two fusion proteins comprising NGF or BDNF fused with elastin-like peptides (ELPs). The aim of this study was to investigate the biological activity of neurotrophin-ELP (N-ELP) fusion proteins via in vitro culture models. NGF and BDNF were cloned in front of an elastin-like polypeptide sequence V40C2. These proteins were expressed in bacteria as inclusion bodies. These fusion proteins underwent solubilization via 8 M urea and purification via inverse transition cycling (ITC). We measured the particle size and the effect of temperature on precipitated particles using dynamic light scattering (DLS). We used western blot analysis to confirm the specificity of NGF-ELP to tropomyosin receptor kinase A (TrkA) antibody and to confirm the specificity of BDNF-ELP to TrkB antibody. PC12 cells w...Continue Reading

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Citations

Jul 7, 2017·Gene Therapy·D A Monfort, P Koria
Oct 25, 2017·Journal of Bioactive and Compatible Polymers·Alex Leonard, Piyush Koria
Dec 24, 2018·Journal of Biomedical Science·Rossana BoniAndrew N Clarkson
Jul 10, 2019·Matrix Biology : Journal of the International Society for Matrix Biology·Arturo Ibáñez-FonsecaJosé Carlos Rodríguez-Cabello
Oct 24, 2020·Advances in Wound Care·Tabitha BoeringerPiyush Koria
Sep 7, 2021·Biotechnology Progress·Graham Strauss, Piyush Koria

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