Henriette Engen Berg

• Berg, Henriette Sjånes; Sæterdal, Kristina Erikstad; Smetop, Tone; Rozenvalds, Rūdolfs; Brandtzaeg, Ole Kristian; Vehus, Tore; Lundanes, Elsa & Wilson, Steven Ray Haakon (2017). Self-packed core shell nano liquid chromatography columns and silica-based monolithic trap columns for targeted proteomics. Journal of Chromatography A.  ISSN 0021-9673.  1498, s 111- 119 . doi: 10.1016/j.chroma.2017.03.043
• Wilson, Steven Ray Haakon; Vehus, Tore; Berg, Henriette Sjånes & Lundanes, Elsa (2015). Nano-LC in proteomics: Recent advances and approaches. Bioanalysis.  ISSN 1757-6180.  7(14), s 1799- 1815 . doi: 10.4155/bio.15.92

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• Amundsen, Sunniva Furre; Wilson, Steven Ray Haakon; Berg, Henriette Engen; Røberg-Larsen, Hanne & Lundanes, Elsa (2018). Online entrapment of epitope-containing peptides from integrins using immobilized antibodies.
• Berg, Henriette Sjånes; Bjørseth, Kamilla; Sæterdal, Kristina Erikstad; Smetop, Tone; Skottvoll, Frøydis Sved; Vehus, Tore; Lundanes, Elsa & Wilson, Steven Ray Haakon (2018). Evaluation of isolation methods of glioma exosomes for biomarker discovery. Show summary

  Gliomas are the most common form of brain cancer, where the sub-group glioblastoma multiforme (GBM) is the most aggressive and the most common in adults [1]. For diagnostic, prognostic, and treatment monitoring, fast and reliable analytical methods for detection of glioblastoma tumors are essential. Non-invasive monitoring of circulating biomarkers in blood (liquid biopsy) is a desirable possibility [2]. Nano LC-MS can provide the high sensitivity, selectivity and low false positive rate needed for the proteomic analysis of exosomes, which are extracellular vesicles released from the tumor and into the bloodstream. We have optimized the packing of 50 µm capillaries (2.6 µm C18 core-shell particles) with peak capacities comparable to similar commercial nano-LC columns (75 µm ID) for peptide separations of minute samples (e.g. exosomes). These columns have previously been successfully used for the targeted detection of CYP27a (a potential breast cancer biomarker) in the MDA-MB-231 cell line. In the present study, these in-house packed columns have been applied for proteins in exosomes isolated from GBM cell culture medium. The aim of this study was to compare isolation methods of GMB exosomes for biomarker discovery and study proteins related to GBM in exosomes. Ultracentrifugation (“golden standard” in exosome isolation) was compared to a Total Exosome Isolation Reagent from Invitrogen (Thermo Fisher Scientific) regarding sample volume, total protein amount, purity, and the presence of exosome markers and GBM biomarker candidates. References 1. Molina, J.R., Y. Hayashi, C. Stephens, and M.M. Georgescu, Invasive Glioblastoma Cells Acquire Stemness and Increased Akt Activation. Neoplasia. 12 (2010) 453-U37. 2. Best, M.G., N. Sol, S. Zijl, J.C. Reijneveld, P. Wesseling, and T. Wurdinger, Liquid biopsies in patients with diffuse glioma. Acta Neuropathologica. 129 (2015) 849-865.

• Hassine, Esma Ben; Berg, Henriette Engen; Brandtzaeg, Ole Kristian; Vehus, Tore; Wilson, Steven Ray Haakon & Lundanes, Elsa (2018). Capillary poly(styrene-co-octadecene-co-divinylbenzene) monolithic columns.
• Hvinden, Ingvild Comfort; Rise, Frode; Berg, Henriette Engen; Lundanes, Elsa & Wilson, Steven Ray Haakon (2018). Approaches to untargeted metabolomics of glioblastoma: NMR and MS.
• Hvinden, Ingvild Comfort; Wilson, Steven Ray Haakon; Rise, Frode; Berg, Henriette Engen & Lundanes, Elsa (2018). Nuclear magnetic resonance spectroscopy based metabolomics discovers biomarkers of glioblastoma drug response. Show summary

  Glioblastoma is the most common and aggressive form of brain cancer. Even with comprehensive treatment regimes, patients face an expected survival of only 15 months. Currently, methods for assessing treatment response are lacking; it is difficult to accurately determine the efficacy of a treatment. The goal of the present study was to contribute to treatment assessment by scouting for metabolic biomarkers occurring in response to exposure to chemotherapeutic agents temozolomide (TMZ) and sepantronium bromide (YM155). Untargeted metabolomics of lysate from cultured glioblastoma cells was carried out with liquid state proton nuclear magnetic resonance (NMR) spectroscopy at resonance frequency 800 MHz. Spectral data were analyzed with two different multivariate statistical methods: principal component analysis (PCA) and partial least squares (PLS) regression. For YM155, two biomarker candidates were found: citric and lactic acid. Citric acid appeared to increase most in samples from cell lines less sensitive to YM155. Lactic acid decreased in all cell lines and was considered a more general biomarker of treatment exposure. TMZ-treated samples were not distinguishable from control samples, most likely due to too short exposure time (24 hours). Analyses with nano hydrophilic interaction liquid chromatography coupled with mass spectrometry (MS) corroborated the findings by NMR spectroscopy and statistical analyses. Both citric acid and lactic acid are biomarker candidates, but a more detailed understanding of their fluctuations in glioblastoma during treatment is needed. Nevertheless, they represent genuine candidates and should be considered for further in vivo magnetic resonance spectroscopy (MRS) studies. In the future, the biomarkers could be monitored with MRS, allowing a more unambiguous and personalized assessment of response to treatment in individual patients.

• Skottvoll, Frøydis Sved; Berg, Henriette Engen; Lund, Kaja; Røberg-Larsen, Hanne; Wilson, Steven Ray Haakon & Lundanes, Elsa (2017). Critical evaluation of isolation and characterization techniques of breast cancer exosomes.
• Sved Skottvoll, Frøydis; Lund, Kaja; Røberg-Larsen, Hanne; Berg, Henriette Sjånes; Thiede, Bernd; Lundanes, Elsa & Wilson, Steven Ray Haakon (2017). Diagnostic potential of exosomes: The purification issue.
• Berg, Henriette Sjånes; Lundanes, Elsa; Wilson, Steven Ray Haakon & Vehus, Tore (2016). Optimization of in-house packing of nano liquid chromatography columns - How to pack low-cost nano liquid chromatography columns?. Show summary

  Liquid chromatography-mass spectrometry (LC-MS) approaches for measurement of biomarkers are increasing in popularity due to the selectivity and sensitivity. To match the detection limits of the immunoassay approaches (such as enzyme-linked immunosorbent assay (ELISA) and Western Blot (WB)), LC columns with narrow inner diameter are preferable. The goal of this work was to establish a simple and low cost slurry packing procedure to obtain 50 µm inner diameter (ID) x 150 mm particle packed nanoLC columns with comparable performance to a 75 µm ID x 150 mm commercial nanoLC column. Slurry packing with 2.6 µm solid-core reversed phase particles was performed with an in-house made pressure bomb system (< 200 bar). Column efficiency was measured as peak capacity (number of peaks separated in a gradient window, PC), calculated using peptides from tryptic digest of human serum albumin (HSA). To maintain sedimentation of particles, heat or magnetic stirring was needed. 80/20 % acetonitrile (ACN)/water as slurry solvent was found to give both the fastest packing and the most efficient columns measured as PC (89 ± 11 at 10 % of the peak height for columns packed with magnetic stirring). Nevertheless, the column performance was quite similar for all slurry solvents and packing approaches. “Pico Frit” column housings from New Objective and two frit-making procedures (polymerization and sintering of silica particles) for standard column housings were compared, but none outperformed the others. The PC of the commercial column packed with the same material as the in-house packed columns, was approximately 30 % higher than the best performing in-house packed column (102 vs 136). However, the retention time repeatability between replicates was equal to the commercial column for most columns packed with 80/20 % ACN/water. In conclusion, this work presents a fast and low cost packing procedure of nanoLC columns with similar performance to commercial nanoLC columns.

• Berg, Henriette Sjånes; Vehus, Tore; Lundanes, Elsa & Wilson, Steven Ray Haakon (2016). Optimization of in-house packed nano-LC columns for peptide separations.

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