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.

公开(公告)号：US20220260584A1

公开(公告)日：2022-08-18

申请号：US17616366

申请日：2020-06-05

Applicant: AMGEN INC.

Inventor： Pavel BONDARENKO ,  Scott Thomas KUHNS ,  Andrew NICHOLS ,  Gang XIAO ,  Liuqing SHI ,  Jill A. CROUSE-ZEINEDDINI

IPC: G01N33/68 ,  G01N33/543 ,  G01N33/547

Abstract: Provided herein are methods of identifying structures, e.g., attributes, of a therapeutic protein or a target that affect an interaction between the therapeutic protein and the target. In exemplary embodiments, the method comprises: (a) applying a stress to a first sample comprising therapeutic proteins or targets; (b) contacting the first sample with a second sample comprising targets or therapeutic proteins to form a mixture comprising (i) therapeutic protein-target complexes, (ii) unbound therapeutic proteins, and (iii) unbound targets; and (c) separating the mixture into at least two fractions, wherein an unbound fraction comprises unbound therapeutic proteins or unbound targets and a bound fraction comprises therapeutic protein-target complexes; and (d) for each of the unbound fraction and bound fraction, identifying and quantifying the abundance of the structures, e.g., attribute, present on a species of the therapeutic protein or target.