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Generation and tumor recognition properties of two human monoclonal antibodies specific to cell surface anionic phospholipids

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Summary

Phosphatidylserine (PS) and other anionic phospholipids, which become exposed on the surface of proliferating endothelial cells, tumor cells and certain leukocytes, have been used as targets for the development of clinical-stage biopharmaceuticals. One of these products (bavituximab) is currently being investigated in Phase 3 clinical trials. There are conflicting reports on the ability of bavituximab and other antibodies to recognize PS directly or through beta-2 glycoprotein 1, a serum protein that is not highly conserved across species. Here, we report on the generation and characterization of two fully human antibodies directed against phosphatidylserine. One of these antibodies (PS72) bound specifically to phosphatidylserine and to phosphatidic acid, but did not recognize other closely related phospholipids, while the other antibody (PS41) also bound to cardiolipin. Both PS72 and PS41 stained 8/9 experimental tumor models in vitro, but both antibodies failed to exhibit a preferential tumor accumulation in vivo, as revealed by quantitative biodistribution analysis. Our findings indicate that anionic phospholipids are exposed and accessible in most tumor types, but cast doubts about the possibility of efficiently targeting tumors in vivo with PS-specific reagents.

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Abbreviations

PS:

Phosphatidylserine

β2GP1:

Beta-2 glycoprotein 1

DPPS:

1,2-Dipalmitoyl-sn-glycero-3-phosphoserine, sodium salt

DPPA:

1,2-Dipalmitoyl-sn-glycero-3-phosphate, sodium salt

DPPC:

1,2-Dipalmitoyl-sn-glycero-3-phosphocholine

DPPE:

1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamine

SM:

Sphingomyelin

CL:

Cardiolipin

PI:

Phosphatidylinositol

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Acknowledgments

Financial support from ETH Zürich, the Swiss National Science Foundation, the Kommission für Technologie und Innovation (KTI MedTech Award) and the European Union (PRIAT FP7 Project) is gratefully acknowledged. Also, we would like to thank Dr. Sarah Wulhfard and Dr. Doriana Triggiani for their help with the biodistribution experiment.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Institute of Pharmaceutical Sciences, Department of Chemistry and Applied Biosciences, ETH Zürich, Vladimir-Prelog-Weg 2, CH-8093, Zurich, Switzerland

    Emil Bujak & Dario Neri

  2. Philochem AG, Libernstrasse 3, CH-8112, Otelfingen, Switzerland

    Francesca Pretto

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  1. Emil Bujak
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  2. Francesca Pretto
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Correspondence to Dario Neri.

Electronic supplementary material

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Supplementary Figure 1

BIAcore profiles of scFv(PS41) and scFv(PS72) against DPPS on L1 BIAcore chip. (JPEG 28 kb)

Supplementary Figure 2

Biodistribution studies of radioiodinated SIP(PS41), SIP(PS72) and SIP(F8) 24 h after i.v. injection into tumor bearing (SKRC-52) BALB/c nude mice. (PPT 187 kb)

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Bujak, E., Pretto, F. & Neri, D. Generation and tumor recognition properties of two human monoclonal antibodies specific to cell surface anionic phospholipids. Invest New Drugs 33, 791–800 (2015). https://doi.org/10.1007/s10637-015-0248-0

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