A streamlined, automated workflow to screen and triage large numbers of baculoviruses for protein expression.

Autor: Kostas JJ; Protein & Structural Chemistry, Discovery Chemistry, Merck & Co., Inc., 770 Sumneytown Pike, West Point, PA 19486, USA., Partridge AT; Protein & Structural Chemistry, Discovery Chemistry, Merck & Co., Inc., 770 Sumneytown Pike, West Point, PA 19486, USA., Byrne NJ; Protein & Structural Chemistry, Discovery Chemistry, Merck & Co., Inc., 770 Sumneytown Pike, West Point, PA 19486, USA., Edwards RW; Evotec (US) Inc., 303B College Road East, Princeton, NJ 08540, USA., Su HP; Protein & Structural Chemistry, Discovery Chemistry, Merck & Co., Inc., 770 Sumneytown Pike, West Point, PA 19486, USA., Gabelli SB; Protein & Structural Chemistry, Discovery Chemistry, Merck & Co., Inc., 770 Sumneytown Pike, West Point, PA 19486, USA., Brooun A; Protein & Structural Chemistry, Discovery Chemistry, Merck & Co., Inc., 770 Sumneytown Pike, West Point, PA 19486, USA., Shipman JM; Protein & Structural Chemistry, Discovery Chemistry, Merck & Co., Inc., 770 Sumneytown Pike, West Point, PA 19486, USA. Electronic address: jennifer_shipman@merck.com.
Jazyk: angličtina
Zdroj: Protein expression and purification [Protein Expr Purif] 2024 Mar; Vol. 215, pp. 106406. Date of Electronic Publication: 2023 Nov 21.
DOI: 10.1016/j.pep.2023.106406
Abstrakt: The baculovirus expression system is a powerful and widely used method to generate large quantities of recombinant protein. However, challenges exist in workflows utilizing either liquid baculovirus stocks or the Titerless Infected-Cells Preservation and Scale-Up (TIPS) method, including the time and effort to generate baculoviruses, screen for protein expression and store large numbers of baculovirus stocks. To mitigate these challenges, we have developed a streamlined, hybrid workflow which utilizes high titer liquid virus stocks for rapid plate-based protein expression screening, followed by a TIPS-based scale-up for larger protein production efforts. Additionally, we have automated each step in this screening workflow using a custom robotic system. With these process improvements, we have significantly reduced the time, effort and resources required to manage large baculovirus generation and expression screening campaigns.
(Copyright © 2023 Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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