Methods for characterizing low-abundance host cell proteins
AssigneeRegeneron Pharmaceuticals, Inc.
InventorsI-Hsuan Chen, Hui Xiao, Ning Li
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Abstract
The present application provides methods and systems to identify host cell protein (HCP) impurities in a sample containing high-abundance proteins. The HCP impurities can be enriched using interacting peptide ligands which have been attached to solid support. The HCP impurities can be eluted from the solid support using solution containing phase transfer surfactants. The isolated HCP impurities can be digested to generate components of the isolated HCP impurities which can subsequently be identified using a mass spectrometer.
Claims20
A method of identifying at least one host cell protein (HCP) impurity in a sample, comprising:
- contacting a sample to a solid support,
- wherein said sample includes at least one high-abundance peptide or protein and at least one HCP impurity,
- wherein interacting peptide ligands have been attached to said solid support, and
- wherein said at least one HCP impurity can bind to said interacting peptide ligands;
- washing said solid support using an elution solution to produce an eluate, wherein said elution solution includes at least one surfactant;
- subjecting said eluate to an enzymatic digestion condition to generate a peptide digest;
- subjecting said peptide digest to acidic conditions to produce an acidified peptide digest;
- centrifuging said acidified peptide digest to produce a surfactant-depleted peptide digest;
- and
- subjecting said surfactant-depleted peptide digest to mass spectrometry analysis to identify said at least one HCP impurity.
A system for identifying at least one host cell protein (HCP) impurity in a sample, comprising:
- a solid support;
- interacting peptide ligands,
- wherein said interacting peptide ligands are attached to said solid support, and
- wherein said at least one HCP impurity can bind to the interacting peptide ligands;
- an elution solution comprising at least one surfactant;
- an enzymatic digestion solution;
- an acidic solution;
- a centrifuge;
- and
- a mass spectrometer capable of identifying said at least one HCP impurity.
Dependencies are inferred from the claim text.