公开(公告)号：WO2022061092A1

公开(公告)日：2022-03-24

申请号：PCT/US2021/050842

申请日：2021-09-17

Applicant: AMGEN INC.

Inventor： XIAO, Gang ,  BONDARENKO, Pavel ,  SHI, Liuqing ,  DILLON, Thomas M. ,  REN, Da ,  RICCI, Margaret Speed ,  CUI, Weidong ,  HARRAHY, John Joseph ,  PAWLOWSKI, Jake ,  DYKSTRA, Andrew ,  RIGGS, Dylan

IPC: C07K14/435 ,  C12N9/76 ,  G01N33/68

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.