title: Quantitation of recombinant protein in whole cells and cell extracts via solid-state NMR spectroscopy.
abstract: Recombinant proteins (RPs) are commonly expressed in bacteria followed by solubilization and chromatography. Purified RP yield can be diminished by losses at any step with very different changes in methods that can improve the yield. Time and labor can therefore be saved by first identifying the specific reason for the low yield. This study describes a new solid-state nuclear magnetic resonance approach to RP quantitation in whole cells or cell extracts without solubilization or purification. The method is straightforward and inexpensive and requires only ∼50 mL culture and a low-field spectrometer.
authors.affiliation: Department of Chemistry, Michigan State University, East Lansing, Michigan 48824, United States.
authors.forename: Erica P
authors.initials: EP
authors.lastname: Vogel
authors.forename: David P
authors.initials: DP
authors.lastname: Weliky
mesh_terms: D001419:Bacteria / Q000737:chemistry / Q000254:growth & development; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D002457:Cell Extracts / Q000737:chemistry; D004274:DNA, Recombinant / Q000737:chemistry; D006358:Hot Temperature; D002479:Inclusion Bodies / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012995:Solubility
chemical_list: D001426:Bacterial Proteins; D002457:Cell Extracts; D004274:DNA, Recombinant; D011994:Recombinant Proteins