Increased production of active human β₂-adrenergic/Gαs fusion receptor in Sf-9 cells using nutrient limiting conditions

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DOIResolve DOI: http://doi.org/10.1006/prep.2001.1476
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TypeArticle
Journal titleProtein Expression and Purification
Volume23
Issue1
Pages6674; # of pages: 9
AbstractUsing the baculovirus/insect-cell expression vector system, we succeeded in obtaining a high yield of active human β₂-adrenergic/Gαs fusion protein. This was achieved following high cell density production under nutrient-limiting conditions using a very low multiplicity of infection (MOI). This approach was found to significantly reduce inactive protein accumulation that occurred when production was done using conventional high MOI procedures. The maximum specific and volumetric yields of active receptor using this strategy increased by factors of two- and sixfold, respectively. Our results suggest that the increase in the ratio of active/total protein produced results from production under nutrient limitation. Since low multiplicity of infection offers many advantages for large-scale applications, we suggest that this simple production method should be considered when optimizing expression of G-protein-coupled receptors and other complex proteins.
Publication date
LanguageEnglish
AffiliationNational Research Council Canada; NRC Biotechnology Research Institute
Peer reviewedNo
NPARC number12328957
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Record identifier5c2cc6e8-1c1e-4bcc-a2e4-818a74a9904d
Record created2009-09-10
Record modified2016-05-09
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