PXD018510

PXD018510 is an original dataset announced via ProteomeXchange.

Title	Hydrogen Deuterium Exchange Mass Spectrometry implicates dynamic CD20 antigen binding to two discontinuous CDR regions in the NCD1.2 monoclonal antibody
Description	A comparative canine-human therapeutics model is being developed in B-cell lymphoma through the generation of a hybridoma cell that produces a murine monoclonal antibody specific for canine CD20. The hybridoma cell produces two light chains, light chain-3 and light chain-7. However, the contribution of either light chain to the authentic full-length hybridoma derived IgG is undefined. Mass spectrometry was used to identify only one of the two light chains, light chain-7, as predominating in the full-length IgG. Gene synthesis created a recombinant murine-canine chimeric monoclonal antibody expressing light chain-7 that reconstituted the IgG binding to CD20. Hydrogen deuterium exchange mass spectrometry was used to define two stages in the mode of binding of the CD20 epitope the antibody. In the early stage of the reaction, the antigen interacted with CDR3 (VH). In the equilibrium stages, stable binding occurred to CDR2 (VH) and CDR2 (VL), without any detectable CDR3 (VH) interactions. These data suggest that CDR3 (VH) functions as a transient antigen docking motif to nucleate the peptide into the antibody active site which resolves into antigen binding with the heavy and light chain CDR2 domains. These approaches define a methodology for fine mapping of CDR contacts using nested enzymatic reactions and hydrogen deuterium exchange mass spectrometry to map the kinetic mode of antigen binding. These data support the further development of an engineered synthetic antibody for use as a canine lymphoma therapeutic that mimics the human anti-CD20 antibody therapeutic.
HostingRepository	PRIDE
AnnounceDate	2022-02-15
AnnouncementXML	Submission_2022-02-15_10:56:07.045.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Lenka Hernychova
SpeciesList	scientific name: Canis lupus x Canis lupus familiaris; NCBI TaxID: 990119;
ModificationList	No PTMs are included in the dataset
Instrument	TripleTOF 5600; LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2020-04-14 01:52:47	ID requested
⏵ 1	2022-02-15 10:56:10	announced

Uhrik L, Hernychova L, Muller P, Kalathiya U, Lisowska MM, Kocikowski M, Parys M, Faktor J, Nekulova M, Nortcliffe C, Zatloukalova P, Ruetgen B, Fahraeus R, Ball KL, Argyle DJ, Vojtesek B, Hupp TR, Hydrogen deuterium exchange mass spectrometry identifies the dominant paratope in CD20 antigen binding to the NCD1.2 monoclonal antibody. Biochem J, 478(1):99-120(2021) [pubmed]

submitter keyword: Human, comparative medicine, CD20, lymphoma, monoclonal antibody, ADEPT, phage display, HDX, LC-MS/MS

Ted Hupp
contact affiliation	University of Edinburgh, Institute of Genetics and Molecular Medicine, Edinburgh, Scotland, United Kingdom, EH4 2XR
contact email	Ted.Hupp@ed.ac.uk
lab head
Lenka Hernychova
contact affiliation	Masaryk Memorial Cancer Institute
contact email	hernychova@seznam.cz
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2021/09/PXD018510

PRIDE project URI

• PRIDE
  1. PXD018510
     1. Label: PRIDE project
     2. Name: Hydrogen Deuterium Exchange Mass Spectrometry implicates dynamic CD20 antigen binding to two discontinuous CDR regions in the NCD1.2 monoclonal antibody