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Pharmacokinetic Analysis of a Novel Human EGFRvIII:CD3 Bispecific Antibody in Plasma and Whole Blood Using a High-Resolution Targeted Mass Spectrometry Approach.

Publication ,  Journal Article
Schaller, TH; Foster, MW; Thompson, JW; Spasojevic, I; Normantaite, D; Moseley, MA; Sanchez-Perez, L; Sampson, JH
Published in: J Proteome Res
August 2, 2019

Bispecific single chain 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-expressing T cells to recognize and destroy malignant, EGFRvIII-expressing glioma. In mice, we showed that EGFRvIII:CD3 bi-scFv effectively treats orthotopic patient-derived malignant glioma and syngeneic glioblastoma. Here, we developed 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 a 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 min and a terminal half-life of ∼2.5 h. Our results establish a sensitive, high-throughput assay for direct quantification of EGFRvIII:CD3 bi-scFv without the need for immunoaffinity enrichment. Moreover, these pharmacokinetic parameters will guide drug optimization and dosing regimens in future IND-enabling and phase I studies of EGFRvIII:CD3 bi-scFv.

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Published In

J Proteome Res

DOI

EISSN

1535-3907

Publication Date

August 2, 2019

Volume

18

Issue

8

Start / End Page

3032 / 3041

Location

United States

Related Subject Headings

  • Xenograft Model Antitumor Assays
  • T-Lymphocytes
  • Proteomics
  • Mice
  • Mass Spectrometry
  • Humans
  • Glioblastoma
  • ErbB Receptors
  • Chromatography, Liquid
  • Cell Line, Tumor
 

Citation

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Schaller, T. H., Foster, M. W., Thompson, J. W., Spasojevic, I., Normantaite, D., Moseley, M. A., … Sampson, J. H. (2019). 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. https://doi.org/10.1021/acs.jproteome.9b00145
Schaller, Teilo H., Matthew W. Foster, J Will Thompson, Ivan Spasojevic, Deimante Normantaite, M Arthur Moseley, Luis Sanchez-Perez, and John H. Sampson. “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, no. 8 (August 2, 2019): 3032–41. https://doi.org/10.1021/acs.jproteome.9b00145.
Schaller TH, Foster MW, Thompson JW, Spasojevic I, Normantaite D, Moseley MA, et al. 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. 2019 Aug 2;18(8):3032–41.
Schaller, Teilo H., et al. “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, vol. 18, no. 8, Aug. 2019, pp. 3032–41. Pubmed, doi:10.1021/acs.jproteome.9b00145.
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. 2019 Aug 2;18(8):3032–3041.
Journal cover image

Published In

J Proteome Res

DOI

EISSN

1535-3907

Publication Date

August 2, 2019

Volume

18

Issue

8

Start / End Page

3032 / 3041

Location

United States

Related Subject Headings

  • Xenograft Model Antitumor Assays
  • T-Lymphocytes
  • Proteomics
  • Mice
  • Mass Spectrometry
  • Humans
  • Glioblastoma
  • ErbB Receptors
  • Chromatography, Liquid
  • Cell Line, Tumor