Assessment of the three-dimensional structure of recombinant protein therapeutics by NMR fingerprinting: demonstration on recombinant human granulocyte macrophage-colony stimulation factor.
We describe a simple, powerful, and robust NMR-based method that has the potential to greatly impact the characterization of recombinant protein therapeutics. The method ascertains the bioactive conformational identity of recombinant human granulocyte macrophage-colony stimulation factor (rhGM-CSF) produced in Streptomyces lividans versus Escherichia coli by overlaying their 2D 1H,15N HSQC correlation spectra. An identical match of all resonances implies that rhGM-CSF from both processes share indistinguishable conformations that correlate with in vitro activity. The result of this method is unique among existing methods. It can detect and quantify the active ingredient. Moreover it provides a complete assessment of the conformation with high sensitivity to minor structural changes.
Aubin
Yves
Y
Gingras
Geneviève
G
Sauvé
Simon
S
D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000032:analysis* / Q000737:chemistry* / Q000627:therapeutic use; D006801:Humans; D021621:Imaging, Three-Dimensional; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry; D048170:Streptomyces lividans / Q000737:chemistry
D011994:Recombinant Proteins; D016178:Granulocyte-Macrophage Colony-Stimulating Factor