ADDovenom: Novel Snakebite Therapy Platform of Unparalleled Efficacy, Safety and Affordability

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Establishment of an insect cell-based platform for production of adenovirus-like particles (ADDomer particles) as a snakebite therapy

Publication . Paiva, Cláudia Cristina Sobreiro; Roldão, António; Lisboa, Cristina

Snakebite envenoming is a neglected tropical disease which causes up to 138 thousand deaths per year. Nowadays, snake antivenom is the only available treatment in the market, originated from horse serum, with limited efficiency and non-reproducible production. This study aimed at implementing and improving ADDomer particles production and purification processes towards the development of a next regeneration ADDomer-based antivenom therapy designed to recognize, interact, and remove snake venom toxics from human body. ADDomer particles were produced intracellularly in insect High Five cells using a baculovirus expression vector system and purified using a three-step purification process (incl. freeze-thaw cycles, centrifugation, size-exclusion and anion exchange chromatography) according to baseline bioprocess. Purified ADDomer particles have a size and morphology similar to those previously reported with a production yield of 19 mg ADDomer particles per L of culture, a purity ˃ 99% and a melting point of ~ 54℃ (as assessed by thermal shift assay). To improve ADDomer particles production, different insect cell lines and culture medium were explored, with High Five cells and Insect XpressTM medium being the most promising combination. To improve ADDomer particles purification, an integrated cell lysis and clarification procedure was devised using 0.2% deviron surfactant (for cell lysis) and a 0.005 m2 BioOptimalTM MF-SL hollow fibber (for concentration and clarification), replacing the freeze-thaw cycles and centrifugation techniques applied in baseline bioprocess. Overall, this work demonstrates the potential of an insect cells-based platform for ADDomer particles production and represents the basis for the development of an ADDomer-based snakebite therapy.

2022-12-05Dissertação de mestrado

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Entidade financiadora

European Commission

Programa de financiamento

H2020

Número da atribuição

899670