HPLC assay for the selective resolution of recombinant proteins

Details HPLC assay for the selective resolution of recombinant proteins HPLC assay for the selective resolution of recombinant proteins

1992-06-23

1994-06-21

Housel, James C.

Chemistry: analytical and immunological testing

Peptide, protein or amino acid

436161, 436172, G01N 2164, G01N 33533, G01N 3368

Patent

active

053227956

ABSTRACT:
Proteins that cannot be resolved by conventional HPLC techniques, including reversed-phase HPLC, can be distinguished, and the quantity of a protein of interest monitored in numerous types of in-process samples, including fermentation and down-stream processing samples, by covalently binding monobromobimane to a protein of interest bearing sulfhydryl groups and chromatographing these in-process samples using reversed-phase HPLC, fluorescence detection and UV absorbance detection.

REFERENCES:
Pande, C. et al. "Characterization of the fluorescent bimane derivative of E. coli initiator transfer RNA". Biochem. Biophys. Res. Commun.; 127(1), pp. 49-55, 1985.
Cotgreave et al. "Methodologies for the application of monobromobimane to the simultaneous analysis of soluble and protein thiol components of biological systems". J. Biochem. Biophys. Methods, 13(4-5), 231-49, 1986.
Svardal et al. "Determination of reduced, oxidized, and protein-bound glutathione in human plasma with precolumn derivatization with monobromobimane and liquid chromatography". Anal. Biochem., 184(2), pp. 338-346, 1990.
P. C. Chinn, et al; Determination of Thiol Proteins Using Monobromobimane Labeling and High-Performance Liquid Chromatographic Analysis: Application to Escherichia coli and Thioredoxin; Analytical Biochemistry 159, pp. 143-149 (1986).

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