Species-Crossreactive scFv Against the Tumor Stroma Marker “Fibroblast Activation Protein” Selected by Phage Display From an Immunized FAP−/− Knock-Out Mouse View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2001-07

AUTHORS

Bodo Brocks, Pilar Garin-Chesa, Eva Behrle, John E. Park, Wolfgang J. Rettig, Klaus Pfizenmaier, Dieter Moosmayer

ABSTRACT

BACKGROUND: Fibroblast activation protein (FAP) is a type II membrane protein expressed on tumor stroma fibroblasts in more than 90% of all carcinomas. FAP serves as a diagnostic marker and is potential therapeutic target for treatment of a wide variety of FAP+ carcinomas. Murine tumor stroma models and FAP-specific antibodies are required to investigate the functional role of FAP in tumor biology and its usefulness for drug targeting. We here describe the development of antibodies with crossreactivity for human (hFAP) and murine FAP (mFAP), which share 89% amino acid identity. MATERIAL AND METHODS: An FAP-/- mouse was sequentially immunized with recombinant murine and human FAP-CD8 fusion proteins. Immunoglobulin cDNA derived from hyperimmune spleen cells was used for the construction of a combinatorial single chain Fv (scFv) library. Phage display selection of FAP-specific scFv was performed on immobilized hFAP followed by selection on cells expressing murine FAP. RESULTS: High-affinity, species-crossreactive, FAP-specific scFv were isolated upon sequential phage display selection. A bivalent derivative (minibody M036) constructed thereof was applied for immunohistochemical analyses and allowed detection of FAP expression on stroma cells of different human carcinomas as well as on murine host stroma in a tumor xenograft model. CONCLUSIONS: MB M036, derived from phage display selected species crossreactive scFv, is suitable for tumor stroma targeting and will be a valuable tool in the analyses of the functional role of FAP in tumor biology as well as in the evaluation of the suitability of FAP for drug targeting. More... »

PAGES

461-469

References to SciGraph publications

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf03401851

DOI

http://dx.doi.org/10.1007/bf03401851

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1074947219

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/11683371


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