Protein Expression in Whole Insect Larvae using baculovirus

R. Balcerzak
Allotropic Tech LLC, United States

Keywords: Baculovirus, Proteins, Enzymes, Acetylcholinesterase (AChE), Recombinant

We describe a novel solution to produce hard to express recombinant proteins, using well-established baculovirus expression system combined with whole insect larvae (Trichoplusia ni). The system is highly efficient, scalable and reproducible, which results in high level target protein expression. Over 300 proteins have been expressed using this system: 1) Tissue heterogeneity of larvae (compared to the homogeneity of cultured cells) can be a significant advantage for difficult to produce proteins due to involvement of larval ions, metabolites, and co-factors required for activity of recombinant proteins. 2) High natural lipid content of larvae facilitates production and recovery of many lipid-associated recombinant proteins, which are not readily produced using other expression systems. 3) Improved post-translational modifications and protein maturation including efficient signal peptide cleavage, N-glycosylation, disulfide bond formation, formation of dimers and protein complexes due to presence of native enzymes and chaperones. 4) Co-expression of two or more viruses in whole insect system is much more efficient than in cultured cells resulting in faster and more efficient study of protein interactions. 5) Emphasis on enzymes produced for the USAMRICD’s chemical defense efforts including AChE, PON1 and BChE