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Background reduction in top-down antibody analysis

A top-down, antibody-based technology, applied in the direction of analyzing materials, particle separation tubes, trajectory-stabilized spectrometers, etc., can solve the problems of increasing the ambiguity of MS2 spectrum analysis and the complexity of wide distribution

Pending Publication Date: 2021-11-09
DH TECH DEVMENT PTE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, a possible obstacle to the "top-down" approach is the broad distribution of the masses of the various multiply charged precursors after ionization and the MS 2 spectral complexity
Furthermore, real-world applications involve complex unknown samples, which further increases the MS 2 Ambiguity in spectral analysis

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  • Background reduction in top-down antibody analysis
  • Background reduction in top-down antibody analysis
  • Background reduction in top-down antibody analysis

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example 1

[0045] A sample including a humanized monoclonal IgG antibody (NIST-mAb) obtained from NIST was prepared. Samples were subjected to desalting and LC separation using a desalted LC column (water), then subjected to electrospray ionization and mass spectrometry using a research-grade quadrupole-TOF system (SCIEX) modified To include an ECD unit as described in the article entitled "Electron Capture Dissociation in a Branched Radio-Frequency Ion Trap" published in Anal. Chem. 87(1):785-792 (which is incorporated by reference in its entirety). The ECD unit is installed between Q1 and Q2 of the mass spectrometer system. A typical e-beam irradiation time is 10 ms, and the e-beam intensity is adjusted for proper dissociation efficiency. TOF-MS has a mass resolution of 35000-47000 and it resolves isotopic patterns of fragments up to Z ~ 30+. Figure 5A Depicts the spectrum produced by an ionized sample where the RF amplitude was chosen to be at 400V p-p allows essentially all ions ...

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Abstract

Method and devices for performing top down analysis of an antibody are described which involve utilizing an ion source to generate a plurality of ions from a sample containing at least one intact antibod. Further, transmitting said plurality of ions through a quadrupole rod set while applying an RF signal thereto and in the absence of a resolving DC voltage so as to preferentially transmit precursor ions having an m / z value greater than a low mass cutoff of about 1500 m / z from the quadrupole rod set to an ECD cell. The method and device can also performing an ECD reaction in the ECD cell on said precursor ions and also detect reaction products from the ECD reaction.

Description

[0001] related application [0002] This application claims the benefit of priority to U.S. Provisional Application No. 62 / 847,130, filed May 13, 2019, which is incorporated herein by reference in its entirety. technical field [0003] The present invention relates to methods and devices utilizing electron capture dissociation (ECD) in the mass spectrometric analysis of intact antibodies. Background technique [0004] Mass spectrometry (MS) is an analytical technique for determining the elemental composition of a test substance, which has both quantitative and qualitative applications. For example, MS can be used to identify unknown compounds and / or determine the structure of a particular compound by observing its fragmentation. Recently, MS has played an increasingly important role in proteomics due to the speed, specificity, and sensitivity of MS strategies in characterizing and identifying peptides and proteins. [0005] One strategy for characterizing proteins in MS-ba...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68H01J49/42
CPCG01N33/6848H01J49/42G01N33/6854H01J49/0054H01J49/4215H01J49/426
Inventor 马场崇P·鲁米恩
Owner DH TECH DEVMENT PTE