gms | German Medical Science

64th Annual Meeting of the German Society of Neurosurgery (DGNC)

German Society of Neurosurgery (DGNC)

26 - 29 May 2013, Düsseldorf

Evaluation of TaqMan® protein assays in meningiomas

Meeting Abstract

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  • Christina Pfister - Klinik für Neurochirurgie, Universitätsklinikum Tübingen
  • Susan Noell - Klinik für Neurochirurgie, Universitätsklinikum Tübingen
  • Florian Roser - Klinik für Neurochirurgie, Universitätsklinikum Tübingen

Deutsche Gesellschaft für Neurochirurgie. 64. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC). Düsseldorf, 26.-29.05.2013. Düsseldorf: German Medical Science GMS Publishing House; 2013. DocP 142

doi: 10.3205/13dgnc559, urn:nbn:de:0183-13dgnc5599

Published: May 21, 2013

© 2013 Pfister et al.
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by-nc-nd/3.0/deed.en). You are free: to Share – to copy, distribute and transmit the work, provided the original author and source are credited.


Outline

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Objective: TaqMan® Protein Assays is an innovative method, which detects proteins through amplification of substitute DNA templates. Therefore TaqMan® Protein Assays use antibodies and proximity ligation for quantitative Real-Time PCR. In this study we evaluated TaqMan® Protein Assays in different human tissues, cancer cell lines and meningiomas.

Method: RNA and proteins from five random meningiomas and the human glioblastoma cell line U373 MG were co-isolated and quantified. RNA and proteins of eight different human tissues (Brain, Cerebral Meninges, Colon, Kidney, Liver, Lung, Pancreas and Placenta) and seven cancer cell lines (A-431, Hl-60, HeLa, HepG2, Jurkat, K-562 and MCF-7) were purchased. Five angiogenesis proteins were determined: VEGFA, VEGFR-2 (KDR), Neuropilin 1 (NRP1), PDGF-B and PDGFR-β. For quantitative Real-Time PCR TaqMan® Gene Expression Assays were used. For TaqMan® Protein Assays biotinylated full length antibodies were labeled with oligonucleotides. Detection limit was compared with Western Blot analysis.

Results: Human brain was used for relative quantification in quantitative Real-Time PCR and TaqMan® Protein Assays. PDGFR-β and KDR expression was not detected in almost all probes (2 of 21) with Western Blot analysis. With TaqMan® Protein Assays PDGFR-β and KDR expression was detected in 18 of 21 protein lysates. VEGFA, PDGF-BB and NRP1 were detected in all probes with TaqMan® Protein Assays, but not in Western Blots. There were differences between RNA and protein levels both with increased protein and RNA level.

Conclusions: TaqMan® Protein Assays allowed the detection and quantification of very low protein expression levels in human tissues, cancer cell lines and meningioma primary cells. Direct correlation of relative RNA quantities and associated protein levels showed in part distinct differences.