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

Protein Expr Purif. 2024 Mar:215:106406. doi: 10.1016/j.pep.2023.106406. Epub 2023 Nov 21.

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.

Keywords: Automation; BEVS; BIIC; Baculovirus expression vector system; Baculovirus-infected insect cell; Expression screening; Insect cells; PASS; Plate-based amplification with screen selection; TIPS; Titerless infected-cells preservation and scale-up.

MeSH terms

  • Baculoviridae* / genetics
  • Baculoviridae* / metabolism
  • Genetic Vectors
  • Recombinant Proteins
  • Triage*
  • Workflow

Substances

  • Recombinant Proteins