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University of Wyoming

UW Technologies Available for Licensing


LICENSED -Technology Disclosure: Various Techs Baculovirus-Insect Cell Expression System - LICENSED


The baculovirus-insect cell expression system is widely used to produce recombinant proteins, including glycoproteins, for various biomedical applications. In this system, a recombinant baculovirus is engineered to encode a protein of interest, and then used as a vector to efficiently deliver the foreign gene to a host. The virus also contributes a strong promoter and transcription complex, which provides high-level expression of the foreign gene at the transcriptional level. The host is usually an established lepidopteran insect cell line, which translates the RNA and can process the resulting proteins in eukaryotic fashion. Generally, the baculovirus-insect cell system is considered a good system for recombinant glycoprotein production due to the eukaryotic nature of the host. However, despite its ability to glycosylate recombinant proteins, this system does not typically yield products with terminally sialylated N-glycans. The fundamental basis for this serious limitation is that insect cells lack several functions needed to produce sialylated N-glycans. This is a problem because the presence or absence of terminal sialic acids can strongly influence glycoprotein behavior in various ways.

For the past several years, we have been working on this problem. One of our approaches has been to incorporate genes encoding mammalian N-glycan processing enzymes into the insect cell lines commonly used as hosts for baculovirus expression vectors. Towards this end, we constructed expression plasmids and developed methods for stable transformation of lepidopteran insect cells. We then used these tools to produce transgenic insect cell lines and extensively characterized their biological and biochemical properties. These efforts yielded transgenic insect cell lines that constitutively express mammalian genes encoding functions required for N-glycoprotein sialylation. That is, these efforts have yielded transgenic insect cell lines with humanized N-glycoprotein processing pathways. These cell lines have normal morphologies, growth properties, and remain competent as hosts for baculovirus expression vectors. Further, the constitutively expressed mammalian transgene products participate in N-glycoprotein biosynthesis and support the production of more authentic products. Thus, these cell lines offer a distinct advantage over unmodified hosts for the baculovirus-mediated production of more authentic recombinant N-glycoproteins.

The University of Wyoming has three technologies available in the Baculovirus-Insect Cell Expression System:

  • 99-001 - Modifying Insect Cell Glycosylation Pathways with Baculovirus Expression    Vectors
  • 04-042 - Production of Human Glycosylated Proteins in Transgenic Insects
  • 04-043 - Post-Translational Modification of Proteins in Transgenic Insects

 

 

 

 

Molecular Biology Professor Don Jarvis works with research associate and UW graduate Jared Aumiller in a University of Wyoming laboratory. (UW Photo)