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公开(公告)号:US20230349912A1
公开(公告)日:2023-11-02
申请号:US18026678
申请日:2021-09-17
Applicant: AMGEN INC.
Inventor: Gang XIAO , Pavel BONDARENKO , Liuqing SHI , Thomas M. DILLON , Da REN , Margaret Speed RICCI , Weidong CUI , John Joseph HARRAHY , Jake PAWLOWSKI , Andrew DYKSTRA , Dylan RIGGS
IPC: G01N33/68
CPC classification number: G01N33/6821 , G01N33/6848 , G01N2333/976
Abstract: Provided herein are methods of processing a polypeptide or protein for analysis, e.g., peptide mapping analysis by mass spectrometry. In exemplary embodiments, the method comprises incubating a digested sample at a mildly acidic pH and/or in the presence of a chaotrope, wherein the digested sample is produced by digesting a polypeptide with a protease to produce a digested sample comprising at least two peptides. In exemplary embodiments, the method comprises digesting the polypeptide with a protease which cleaves C-terminal to tryptophan to produce a digested sample comprising at least two peptides. In exemplary embodiments, the method comprises digesting the polypeptide with trypsin at an enzyme:substrate (E:S) weight ratio of about 1:1 to about 1:15 to produce a digested sample comprising at least two peptides. In exemplary aspects, the digested sample comprises at least one or two peptides each comprising a tyrosine at the C-terminus.