InsectDirect™ System

InsectDirect Home   •  How does use of the       InsectDirect System benefit me?   •  What does the InsectDirect System offer?      •  Insect Cell Expression Vectors      •  Insect GeneJuice® Transfection Reagent      •  Insect RoboPop™         Ni-NTA His•Bind® Purification Kit         •  A brief protocol for the RoboPop kit         •  Comparison of traditional baculovirus             method and InsectDirect System method         •  Small- and large-scale Sf9 suspension             culture transfections

How does use of the InsectDirect System benefit me?

• Allows protein expression in 48 hours, not 2 to 3 weeks
• Omits the need for a time-consuming production of recombinant baculovirus
• Provides an option for HT
  screening of multiple targets
• Produces up to 80 µg target
  protein per 1 ml culture
• Includes a protocol that can
  be scaled for larger culture
  volumes and higher protein yields

	A. Traditional baculovirus method			B. InsectDirect System method
	Day 1	Cotransfect insect cells with recombinant transfer plasmid plus linearized AcNPV vector DNA		Day 1	Transfect Sf9 cells with pIEx recombinant plasmid using Insect GeneJuice Transfection Reagent
	Day 4	Choose plaque and replate		Day 3	Proceed with purification
	Day 8	Choose plaque and amplify
	Day 11	Screen for expression; amplify
	Day 14	Titer viral stock; optimize MOI
	Day 1 8	Infect insect cells and express protein
	Days 20–21	Harvest cells and proceed with purification

Researchers dealing with high-throughput (HT) expression screening face the fundamental challenge of trying to rapidly identify the appropriate expression system for many targets in parallel. Often known or unknown open reading frames (ORFs) are amplified by PCR, cloned into a variety of vectors, and the recombinants used to direct target protein expression in E. coli, mammalian cells, insect cells, or yeast. The focus has traditionally not involved insect expression systems due to their complexity and time-consuming protocols. However, insect cell systems provide an alternative means for expressing proteins that require specific post-translational modifications to retain solubility and activity.

To facilitate rapid expression and purification in Spodoptera insect cells (Sf9, Sf21), Novagen developed transient expression vectors, thereby eliminating the need to create recombinant baculovirus for protein expression, with some (vectors) that are a part of our Radiance™ System featuring the HT-compatible Ligation-independent Cloning format (Ek/LIC) . In addition, a high-efficiency insect cell transfection reagent was developed as well as an automation-compatible His•Tag® fusion protein purification system for insect cells to facilitate expression screening.