Methods for characterizing host-cell proteins
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsSook Yen E, Yunli Hu, Rosalynn Molden, Haibo Qiu, Ning Li
Life of this patent
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Abstract
A practical and effective method for low abundance host cell protein (HCP) identification and quantification, which facilitates the downstream purification process in eliminating potentially problematic HCPs. In particular, the method comprises performing a native digestion followed by characterization using Multiple Reaction Monitoring approach.
Claims20
A method for identifying at least one host-cell protein in a sample matrix including a protein of interest, comprising:
- (a) adding a digestion agent under native conditions to obtain a native digested sample;
- and
- (b) analyzing the native digested sample using a triple quadrupole mass spectrometer to identify a peptide specific to said at least one host-cell protein, wherein said triple quadrupole mass spectrometer is run to obtain multiple reaction monitoring for at least one precursor-product ion for said peptide.
A method for quantifying at least one host-cell protein in a sample matrix including a protein of interest, comprising:
- (a) adding a digestion agent under native conditions to obtain a native digested sample;
- (b) analyzing the native digested sample using a triple quadrupole mass spectrometer to identify a peptide specific to said at least one host-cell protein, wherein said triple quadrupole mass spectrometer is run to obtain multiple reaction monitoring for at least one precursor-product ion for said peptide;
- (c) quantifying an amount of said at least one host-cell protein by quantifying an amount of said peptide by utilizing an isotopically-enriched peptide having the same amino acid sequence as that of said peptide to calibrate the quantitation of said peptide, and utilizing a standard comprising a known concentration of peptide to calibrate the quantitation of said peptide.
Dependencies are inferred from the claim text.