Uliana J Voytovich

Publications (3)36.61 Total impact

• Article: Multiplexed quantification of estrogen receptor and HER2/Neu in tissue and cell lysates by peptide immunoaffinity enrichment mass spectrometry.

  Regine M Schoenherr, Jeffrey R Whiteaker, Lei Zhao, Richard G Ivey, Mary Trute, Jacob Kennedy, Uliana J Voytovich, Ping Yan, Chenwei Lin, Amanda G Paulovich
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  ABSTRACT: Access to a wider range of quantitative protein assays would significantly impact the number and use of tissue markers in guiding disease treatment. Quantitative mass spectrometry-based peptide and protein assays, such as immuno-SRM assays, have seen tremendous growth in recent years in application to protein quantification in biological fluids such as plasma or urine. Here, we extend the capability of the technique by demonstrating the application of a multiplexed immuno-SRM assay for quantification of estrogen receptor (ER) and human epidermal growth factor receptor 2 (HER2) levels in cell line lysates and human surgical specimens. The performance of the assay was characterized using peptide response curves, with linear ranges covering approximately four orders of magnitude and limits of detection in the low fmol/mg lysate range. Reproducibility was acceptable with median coefficients of variation of approximately 10%. We applied the assay to measurements of ER and HER2 in well-characterized cell line lysates with good discernment based on ER/HER2 status. Finally, the proteins were measured in surgically resected breast cancers, and the results showed good correlation with ER/HER2 status determined by clinical assays. This is the first implementation of the peptide-based immuno-SRM assay technology in cell lysates and human surgical specimens.
  Proteomics 04/2012; 12(8):1253-60. · 4.43 Impact Factor
• Source

  Available from: Karen S Kelly-Spratt

  Article: A targeted proteomics-based pipeline for verification of biomarkers in plasma.

  Jeffrey R Whiteaker, Chenwei Lin, Jacob Kennedy, Liming Hou, Mary Trute, Izabela Sokal, Ping Yan, Regine M Schoenherr, Lei Zhao, Uliana J Voytovich, [......], Philip R Gafken, Jason M Hogan, Lisa A Jones, Pei Wang, Lynn Amon, Lewis A Chodosh, Peter S Nelson, Martin W McIntosh, Christopher J Kemp, Amanda G Paulovich
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  ABSTRACT: High-throughput technologies can now identify hundreds of candidate protein biomarkers for any disease with relative ease. However, because there are no assays for the majority of proteins and de novo immunoassay development is prohibitively expensive, few candidate biomarkers are tested in clinical studies. We tested whether the analytical performance of a biomarker identification pipeline based on targeted mass spectrometry would be sufficient for data-dependent prioritization of candidate biomarkers, de novo development of assays and multiplexed biomarker verification. We used a data-dependent triage process to prioritize a subset of putative plasma biomarkers from >1,000 candidates previously identified using a mouse model of breast cancer. Eighty-eight novel quantitative assays based on selected reaction monitoring mass spectrometry were developed, multiplexed and evaluated in 80 plasma samples. Thirty-six proteins were verified as being elevated in the plasma of tumor-bearing animals. The analytical performance of this pipeline suggests that it should support the use of an analogous approach with human samples.
  Nature Biotechnology 06/2011; 29(7):625-34. · 29.50 Impact Factor
• Article: Blood-based detection of radiation exposure in humans based on novel phospho-Smc1 ELISA.

  Richard G Ivey, Heather D Moore, Uliana J Voytovich, Cortlandt P Thienes, Travis D Lorentzen, Era L Pogosova-Agadjanyan, Shani Frayo, Venissa K Izaguirre, Sally J Lundberg, Lacey Hedin, Kas Ray Badiozamani, Andrew N Hoofnagle, Derek L Stirewalt, Pei Wang, George E Georges, Ajay K Gopal, Amanda G Paulovich
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  ABSTRACT: The structural maintenance of chromosome 1 (Smc1) protein is a member of the highly conserved cohesin complex and is involved in sister chromatid cohesion. In response to ionizing radiation, Smc1 is phosphorylated at two sites, Ser-957 and Ser-966, and these phosphorylation events are dependent on the ATM protein kinase. In this study, we describe the generation of two novel ELISAs for quantifying phospho-Smc1(Ser-957) and phospho-Smc1(Ser-966). Using these novel assays, we quantify the kinetic and biodosimetric responses of human cells of hematological origin, including immortalized cells, as well as both quiescent and cycling primary human PBMC. Additionally, we demonstrate a robust in vivo response for phospho-Smc1(Ser-957) and phospho-Smc1(Ser-966) in lymphocytes of human patients after therapeutic exposure to ionizing radiation, including total-body irradiation, partial-body irradiation, and internal exposure to (131)I. These assays are useful for quantifying the DNA damage response in experimental systems and potentially for the identification of individuals exposed to radiation after a radiological incident.
  Radiation Research 03/2011; 175(3):266-81. · 2.68 Impact Factor