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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.

SUBMITTER: Vogel EP

PROVIDER: S-EPMC3734946 | biostudies-literature | 2013 Jun

REPOSITORIES: biostudies-literature

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Quantitation of recombinant protein in whole cells and cell extracts via solid-state NMR spectroscopy.

Vogel Erica P EP Weliky David P DP

Biochemistry 20130617 25

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. Th ...[more]

PMID: 23742073

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