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A streamlined, automated workflow to screen and triage large numbers of baculoviruses for protein expression.
Kostas, James J; Partridge, Andrea T; Byrne, Noel J; Edwards, Richard W; Su, Hua-Poo; Gabelli, Sandra B; Brooun, Alexei; Shipman, Jennifer M.
Afiliação
  • 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.
Protein Expr Purif ; 215: 106406, 2024 Mar.
Article em En | MEDLINE | ID: mdl-37995943
ABSTRACT
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.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Triagem / Baculoviridae Idioma: En Ano de publicação: 2024 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Triagem / Baculoviridae Idioma: En Ano de publicação: 2024 Tipo de documento: Article