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Clinical Chemistry 54: 597-601, 2008; 10.1373/clinchem.2007.096099
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(Clinical Chemistry. 2008;54:597-601.)
© 2008 American Association for Clinical Chemistry, Inc.

Brief Communications

A New ELISA for Use in a 3-ELISA System to Assess Concentrations of VEGF Splice Variants and VEGF110 in Ovarian Cancer Tumors

Johnny Gutierrez1, Gottfried E. Konecny2, Kyu Hong1, Alexander Burges3, Timothy D. Henry4, Pier D. Lambiase5, Wai Lee Wong1 and Y. Gloria Meng1,a

1 Department of Assay and Automation Technology, Genentech Inc., South San Francisco, CA; 2 Division of Hematology-Oncology, Department of Medicine, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA;3 Department of Obstetrics and Gynecology, Klinikum Grosshadern, Ludwig Maximilians Universität München, Munich, Germany; 4 Division of Cardiology at Hennepin County Medical Center, Minneapolis, MN; 5 Department of Cardiology, Rayne Institute, St Thomas’ Hospital, London, UK;

aaddress correspondence to this author at: Department of Assay and Automation Technology, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080. Fax 650-225-1770; e-mail meng.gloria{at}gene.com.


Abstract

Background: Vascular endothelial growth factor (VEGF), which affects tumor angiogenesis, is expressed as different splice variants, including the major isoforms VEGF165 and VEGF121, and can be cleaved by plasmin to generate VEGF110. The amount of VEGF121 and VEGF110 in biological samples has not been well studied.

Methods: We developed an ELISA that detects VEGF165 and VEGF121 equally, but does not detect VEGF110. We used this ELISA together with 2 other ELISAs, one detecting VEGF165 and the other detecting VEGF165, VEGF121, and VEGF110 equally, to assess the concentrations of VEGF121 and VEGF110 in ovarian cancer tumors.

Results: The median concentrations in ovarian cancer tumor lysates were 0.61 (range <0.055–74) fmol/mg protein for VEGF165, 1.4 (range <0.20–500) fmol/mg protein for VEGF165 plus VEGF121, and 2.3 (range <0.079–520) fmol/mg protein for total VEGF including VEGF110 (n = 248). VEGF concentrations measured by the 3 ELISAs were highly correlated (r = 0.91–0.94). Median estimated VEGF121 and VEGF110 concentrations were 0.77 and 0.58 fmol/mg protein, respectively. In lysates with measurable VEGF165 and total VEGF concentrations, mean VEGF165 was approximately 31% (SD 23%) of the total VEGF (n = 217). In contrast, VEGF165 constituted approximately half of the total circulating VEGF.

Conclusion: VEGF165, VEGF121, and VEGF110 may be present at significant amounts in ovarian cancer tumors.






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Copyright © 2008 by the American Association for Clinical Chemistry.