Production of selenomethionyl-derivatized proteins in baculovirus-infected insect cells.
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 approximately 75% and with a recovery of approximately 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 Sf9 or Trichoplusia ni High Five insect cells as the expression host, and when cultures are grown in either shake flasks or in Wave BioReactors.
Cronin
Ciarán N
CN
Pfizer, Inc., 10777 Science Center Drive, San Diego, CA 92121, USA. ciaran.cronin@pfizer.com
Lim
Kheng B
KB
Rogers
Joe
J
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D019149:Bioreactors; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D003470:Culture Media; D009036:Moths / Q000166:cytology* / Q000821:virology; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012645:Selenomethionine / Q000378:metabolism*; D018411:Spodoptera / Q000166:cytology* / Q000821:virology; D014764:Viral Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D003470:Culture Media; D011994:Recombinant Proteins; D014764:Viral Proteins; D012645:Selenomethionine