Method for simultaneous quantification of ALXN1210 and eculizumab in human serum or urine
AssigneeAlexion Pharmaceuticals, Inc.
InventorsMeng Chen, Ryan Pelto
Life of this patent
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Abstract
Provided herein are methods for simultaneously detecting and quantifying antibodies which bind to the same target and have high sequence identity, such as eculizumab and ALXN1210, present together or alone in a biological sample.
Claims28
A method of detecting and quantifying the respective amounts of two antibodies having high sequence identity present together in a biological sample, wherein the antibodies are eculizumab and ravulizumab, the method comprising:
- (a) treating the biological sample containing the antibodies with a protease to form a proteolytic peptide mixture of the antibodies in the biological sample;
- (b) analyzing a sample of the proteolytic peptide mixture by high performance liquid chromatography (HPLC) tandem mass spectrometry to detect a signature peptide from each of the antibodies in the biological sample, wherein the signature peptide for eculizumab comprises SEQ ID NO:
- 1, SEQ ID NO:
- 19, or SEQ ID NO:
- 20, and the signature peptide for ravulizumab comprises SEQ ID NO:
- 2, SEQ ID NO:
- 21, or SEQ ID NO:22 wherein the signature peptide comprises no more than 27 amino acids;
- and
- (c) quantifying each antibody in the biological sample based on the signal ratio of its signature peptide to an internal control, wherein the internal control comprises a labeled form of the same signature peptide.
A method of detecting and quantifying the amount of an antibody present in a biological sample, wherein the antibody is eculizumab, the method comprising:
- (a) treating the biological sample containing the antibody with a protease to form a proteolytic peptide mixture of the antibody in the biological sample;
- (b) analyzing a sample of the proteolytic peptide mixture by high performance liquid chromatography (HPLC) tandem mass spectrometry to detect a signature peptide, wherein the signature peptide comprises SEQ ID NO:
- 1, SEQ ID NO:
- 19, or SEQ ID NO:
- 20 and wherein the signature peptide comprises no more than 27 amino acids;
- and
- (c) quantifying the amount of eculizumab in the biological sample based on the signal ratio of the signature peptide to an internal control, wherein the internal control comprises a labeled form of the same signature peptide.
A method of detecting and quantifying the amount of an antibody present in a biological sample, wherein the antibody is ravulizumab, the method comprising:
- (a) treating the biological sample containing the antibody with a protease to form a proteolytic peptide mixture of the antibody in the biological sample;
- (b) analyzing a sample of the proteolytic peptide mixture by high performance liquid chromatography (HPLC) tandem mass spectrometry to detect a signature peptide, wherein the signature peptide comprises SEQ ID NO:
- 2, SEQ ID NO:
- 21, or SEQ ID NO:22 wherein the signature peptide comprises no more than 27 amino acids;
- and
- (c) quantifying the amount of ravulizumab in the biological sample based on the signal ratio of the signature peptide to an internal control, wherein the internal control comprises a labeled form of the same signature peptide.
An isolated peptide consisting of SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 19, SEQ ID NO:
- 21, or SEQ ID NO:
- 22.
Dependencies are inferred from the claim text.