Production and characterization of monoclonal anti-sphingosine-1-phosphate antibodies

Nicole O'Brien, S. Tarran Jones, David G. Williams, H. Brad Cunningham, Kelli Moreno, Barbara Visentin, Angela Gentile, John Vekich, William Shestowsky, Masao Hiraiwa, Rosalia Matteo, Amy Cavalli, Douglas Grotjahn, Maria Grant, Geneviève Hansen, Mary Ann Campbell, Roger Sabbadini

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Abstract

Sphingosine-1-phosphate (S1P) is a pleiotropic bioactive lipid involved in multiple physiological processes. Importantly, dysregulated S1P levels are associated with several pathologies, including cardiovascular and inflammatory diseases and cancer. This report describes the successful production and characterization of a murine monoclonal antibody, LT1002, directed against S1P, using novel immunization and screening methods applied to bioactive lipids. We also report the successful generation of LT1009, the humanized variant of LT1002, for potential clinical use. Both LT1002 and LT1009 have high affinity and specificity for S1P and do not cross-react with structurally related lipids. Using an in vitro bioassay, LT1002 and LT1009 were effective in blocking S1P-mediated release of the pro-angiogenic and prometastatic cytokine, interleukin-8, from human ovarian carcinoma cells, showing that both antibodies can outcompete S1P receptors in binding to S1P. In vivo anti-angiogenic activity of all antibody variants was demonstrated using the murine choroidal neovascularization model. Importantly, intravenous administration of the antibodies showed a marked effect on lymphocyte trafficking. The resulting lead candidate, LT1009, has been formulated for Phase 1 clinical trials in cancer and age-related macular degeneration. The anti-S1P antibody shows promise as a novel, first-in-class therapeutic acting as a "molecular sponge" to selectively deplete S1P from blood and other compartments where pathological S1P levels have been implicated in disease progression or in disorders where immune modulation may be beneficial.

Original languageEnglish (US)
Pages (from-to)2245-2257
Number of pages13
JournalJournal of Lipid Research
Volume50
Issue number11
DOIs
StatePublished - Nov 2 2009

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Keywords

  • Age related macular degeneration
  • Angiogenesis
  • Bioactive lipids
  • Cancer
  • Choroidal neovascularization
  • Complementarity determining region
  • Humanization
  • Lymphocyte trafficking
  • Monoclonal antibodies
  • Sphingosine-1-phosphate

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

Cite this

O'Brien, N., Jones, S. T., Williams, D. G., Cunningham, H. B., Moreno, K., Visentin, B., Gentile, A., Vekich, J., Shestowsky, W., Hiraiwa, M., Matteo, R., Cavalli, A., Grotjahn, D., Grant, M., Hansen, G., Campbell, M. A., & Sabbadini, R. (2009). Production and characterization of monoclonal anti-sphingosine-1-phosphate antibodies. Journal of Lipid Research, 50(11), 2245-2257. https://doi.org/10.1194/jlr.M900048-JLR200