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Production of selenomethionyl‐derivatized proteins in baculovirus‐infected insect cells
Author(s) -
Cronin Ciarán N.,
Lim Kheng B.,
Rogers Joe
Publication year - 2007
Publication title -
protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.353
H-Index - 175
eISSN - 1469-896X
pISSN - 0961-8368
DOI - 10.1110/ps.072931407
Subject(s) - spodoptera , trichoplusia , baculoviridae , recombinant dna , sf9 , biology , laboratory flask , bioreactor , insect , cell culture , microbiology and biotechnology , biochemistry , chemistry , botany , noctuidae , gene , pest analysis , genetics
A protocol is described for the production of both intracellularly expressed and secreted selenomethionyl‐derivatized recombinant proteins in baculovirus‐infected insect cells. The method results in the production of recombinant soluble proteins with an SeMet occupancy of ∼75% and with a recovery of ∼20% that of native protein expression. The method is independent of the percentage methionine content of the protein and is reliable and consistent. Similar results are obtained using either Spodoptera frugiperda Sf 9 or Trichoplusia ni High Five insect cells as the expression host, and when cultures are grown in either shake flasks or in Wave BioReactors.