PXD012472-1

PXD012472 is an original dataset announced via ProteomeXchange.

Title	Pharmacokinetic analysis of a novel human EGFRvIII:CD3 bispecific antibody in plasma and whole blood using a high-resolution targeted mass spectroscopy approach
Description	Bi-specific single change antibody fragments (bi-scFv) represent an emerging class of biotherapeutics. We recently developed a fully human bi-scFv (EGFRvIII:CD3 bi-scFv) with the goal of redirecting CD3-expressiong T cells to recognize and destroy malignant, EGFR-expressing glioma. In mice, we showed that EGFRvIII:CD3 bi-scFv effectively treats orthotopic patient-derived malignant glioma and syngeneic glioblastoma. Here, we sought to develop a targeted assay for pharmacokinetic (PK) analysis of EGFRvIII:CD3 bi-scFv, a necessary step in the drug development process. Using microflow liquid chromatography coupled to high resolution parallel reaction monitoring mass spectrometry, and data analysis in Skyline, we developed a bottom-up proteomic assay for quantification of EGFRvIII:CD3 bi-scFv in both plasma and whole blood. Importantly, a protein calibrator, along with stable isotope-labeled EGFRvIII:CD3 bi-scFv protein, were used for absolute quantification. A PK analysis in a CD3 humanized mouse revealed that EGFRvIII:CD3 bi-scFv in plasma and whole blood has an initial half-life of ~8 minutes and a terminal half-life of ~2.5 hours. Our results establish a sensitive, high-throughput assay for direct quantification of EGFRvIII:CD3 bi-scFv without the need for immunoaffinity enrichment. Moreover, these half-life measurements will guide drug optimization and dosing regimens in future pre-clinical and Phase 0/I studies of EGFRvIII:CD3 bi-scFv
HostingRepository	PanoramaPublic
AnnounceDate	2019-07-09
AnnouncementXML	Submission_2019-07-09_15:00:00.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Matt Foster
SpeciesList	scientific name: Mus musculus; NCBI TaxID: 10090; scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	Carbamidomethyl; Label:13C(6)15N(2); Label:13C(6)15N(4)
Instrument	Q Exactive HF-X

Revision	Datetime	Status	ChangeLog Entry
0	2019-01-22 11:55:58	ID requested
⏵ 1	2019-07-09 15:00:01	announced

Schaller TH, Foster MW, Thompson JW, Spasojevic I, Normantaite D, Moseley MA, Sanchez-Perez L, Sampson JH, Pharmacokinetic Analysis of a Novel Human EGFRvIII:CD3 Bispecific Antibody in Plasma and Whole Blood Using a High-Resolution Targeted Mass Spectrometry Approach. J Proteome Res, 18(8):3032-3041(2019) [pubmed]

submitter keyword: PRM, pharmacokinetics, biotherapeutic antibody, stable isotope-labeled protein, whole blood, HF-X

Matt Foster
contact affiliation	Duke Proteomics and Metabolomics Shared Resource
contact email	mwfoster@duke.edu
lab head
Matt Foster
contact affiliation	Duke Proteomics and Metabolomics Shared Resource
contact email	mwfoster@duke.edu
dataset submitter

Panorama Public dataset URI