Choice of fusion proteins, expression host, and analytics solves difficult-to-produce protein challenges in discovery research.

Autor: Lyons-Abbott S; Novo Nordisk Research Center Seattle, Inc, Seattle, Washington, USA., Abramov A; Novo Nordisk Research Center Seattle, Inc, Seattle, Washington, USA., Chan CL; Novo Nordisk Research Center Seattle, Inc, Seattle, Washington, USA., Deer JR; Novo Nordisk Research Center Seattle, Inc, Seattle, Washington, USA., Fu G; Novo Nordisk Research Center Seattle, Inc, Seattle, Washington, USA., Hassouneh W; Novo Nordisk Research Center Seattle, Inc, Seattle, Washington, USA., Koch T; Novo Nordisk Research Center Seattle, Inc, Seattle, Washington, USA., Misquith A; Novo Nordisk Research Center Seattle, Inc, Seattle, Washington, USA., O'Neill J; Novo Nordisk Research Center Seattle, Inc, Seattle, Washington, USA., Simon SA; Novo Nordisk Research Center Seattle, Inc, Seattle, Washington, USA., Wolf A; Novo Nordisk Research Center Seattle, Inc, Seattle, Washington, USA., Yeh R; Novo Nordisk Research Center Seattle, Inc, Seattle, Washington, USA., Vernet E; Novo Nordisk Research Center Seattle, Inc, Seattle, Washington, USA.
Jazyk: angličtina
Zdroj: Biotechnology journal [Biotechnol J] 2024 Jan; Vol. 19 (1), pp. e2300162. Date of Electronic Publication: 2023 Oct 15.
DOI: 10.1002/biot.202300162
Abstrakt: High quality biological reagents are a prerequisite for pharmacological research. Herein a protein production screening approach, including quality assessment methods, for protein-based discovery research is presented. Trends from 2895 expression constructs representing 253 proteins screened in mammalian and bacterial hosts-91% of which are successfully expressed and purified-are discussed. Mammalian expression combined with the use of solubility-promoting fusion proteins is deemed suitable for most targets. Furthermore, cases utilizing stable cell line generation and choice of fusion protein for higher yield and quality of difficult-to-produce proteins (Leucine-rich repeat-containing G-protein coupled receptor 4 (LGR4) and Neurturin) are presented and discussed. In the case of Neurturin, choice of fusion protein impacted the target binding 80-fold. These results highlight the need for exploration of construct designs and careful Quality Control (QC) of difficult-to-produce protein reagents.
(© 2023 Wiley-VCH GmbH.)
Databáze: MEDLINE
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