Development of a cross-reactive monoclonal antibody for detecting the tumor stroma

Hallie M. Hintz, Aidan E. Cowan, Mariya Shapovalova, Aaron LeBeau

Research output: Contribution to journalArticle

Abstract

Here, we document the discovery of a monoclonal antibody that selectively binds to both human and murine fibroblast activation protein alpha (FAP), a serine protease that is overexpressed on cancer-associated fibroblasts (CAFs), making it an attractive therapeutic target for the aiding and abetting tumor microenvironment. The lead antibody, B12, was identified from a naïve murine single-chain variable fragment antibody phage display library screened against recombinant human FAP on magnetic beads. The heavy and light chains of B12 were cloned into full-length human immunoglobulin 1 (IgG) vectors and expressed as a chimeric monoclonal antibody (B12 IgG). We engineered a drug-resistant prostate cancer cell line, CWR-R1-EnzR, to express human FAP for antibody characterization and validation (R1-EnzRFAP). B12 IgG selectively bound to the R1-EnzRFAP cells by flow cytometry and was internalized in vitro by confocal microscopy. B12 IgG was further evaluated as a near-infrared (NIR) optical imaging probe in R1-EnzRFAP and parental xenograft models. High tumor uptake and retention of the NIR probe was observed in the R1-EnzRFAP xenografts, and endogenous expression of murine stromal origin FAP was detected in the parental xenografts. Ex vivo evaluation of these models by immunohistochemistry documented B12 IgG localization to both human and murine FAP-expressing cells.

Original languageEnglish (US)
Pages (from-to)1466-1476
Number of pages11
JournalBioconjugate Chemistry
Volume30
Issue number5
DOIs
StatePublished - May 15 2019

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Monoclonal antibodies
Fibroblasts
Tumors
Immunoglobulin G
Monoclonal Antibodies
Chemical activation
Heterografts
Proteins
Antibodies
Neoplasms
Infrared radiation
Single-Chain Antibodies
Bacteriophages
Tumor Microenvironment
Flow cytometry
Confocal microscopy
Optical Imaging
Serine Proteases
Confocal Microscopy
Libraries

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Development of a cross-reactive monoclonal antibody for detecting the tumor stroma. / Hintz, Hallie M.; Cowan, Aidan E.; Shapovalova, Mariya; LeBeau, Aaron.

In: Bioconjugate Chemistry, Vol. 30, No. 5, 15.05.2019, p. 1466-1476.

Research output: Contribution to journalArticle

Hintz, Hallie M. ; Cowan, Aidan E. ; Shapovalova, Mariya ; LeBeau, Aaron. / Development of a cross-reactive monoclonal antibody for detecting the tumor stroma. In: Bioconjugate Chemistry. 2019 ; Vol. 30, No. 5. pp. 1466-1476.
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