ESR3: Characterisation of the cell surface glycome on the dopamine neuron using glycochip technology
Michel Riese (University of Manchester)
Supervisor: Sabine Flitsch

Project summary
The project involves the development of a glycoarray platform for the glycan-based biomarker discovery linked to Parkinson’s disease (PD). The combination of MALDI-ToF mass spectrometry and the gold-chip array technology developed in the Flitsch are optimised for the screening of carbohydrate-binding proteins (WP8, D8.5), protein extracts from cell culture (WP8, D8.4), as well as PD patients samples (WP8, D8.1). Appropriate adaptations to the self-assembled monolayer (SAM) glycoarray technology are explored to include the possibility glycolipids analysis (cerebrosides). With the help of proteolytic digestion followed by MS/MS the identification of binding partners will be investigated.

The specific aims of the project are:

  1. MALDI-TOF assay development
  2. Glycoarray analysis of carbohydrate-binding proteins
  3. Glycoarray analysis of SH-SY5Y cells
  4. Glycochip screening of samples

Outputs [top]
In progress.

Training [top]

(i) Local level
'Diversity in the Workplace' Training, 15/05/2014, UMAN This equality and diversity online training was suggested by the University of Manchester (UMAN) to encourage (international) students, researchers and staff members to learn and think about basic human rights, workplace etiquette and team work.

'Careers for PhDs' workshop, 12/02/2015, UMAN Especially tailored for first- and early second-year PhD students, this UMAN workshop focusses on stimulating early thoughts on career planning and management. Basics in idea pitching, interviews, and networking for researchers are discussed.

'Creating Effective Conference Presentations', 27/05/2015, UMAN This hands-on workshop the goal was to create and deliver an effective mini-conference presentation with a focus on both design and delivery. Participants were trained in layout and ‘story- telling’ as well as basic presentation techniques.

'IBCarb' workshop on carbohydrate modelling, 06/07/2015, UMAN A one-day workshop on molecular modelling and structure determination of glycans with lectures given by world-leading expert from the field.

(ii) Network level

NMR Boot camp/Microscopy training (November 2014) This four-day workshop covered a brief introduction into nuclear magnetic resonance spectroscopy (NMR) and applications in the field of biology (functional and conformational studies on peptides and nucleic acids, metabolomics in patient’s samples). Furthermore, Agilent and Trinity College Dublin (TCD) staff provided a practical training on sample preparation and instrumentation including basic experiment setup. The second half was dedicated to fluorescence imaging and microscopy. Following a seminar on the basics of histology, fluorescence and instrumentation, TCD core facility staff ran demonstrations and initial experiments on the range of fluorescence microscopes and flow cytometers.

ANDOR Academy/Seahorse & Oroborus training (May 2015)
The first half of the second TINTIN workshop held at TCD focussed on metabolic analysis of cells and organelles. An introductory seminar covered the biochemical basics in the field of bioenergetics before the two techniques of Seahorse (XF Analyzer) and Oroborus were compared in their respective features. Demonstrations and experiments on these instruments were run by the ESRs themselves under the guidance of TCD researchers. Building on the microscopic knowledge laid out six months earlier, ANDOR ran a comprehensive workshop from their ANDOR Academy series introducing physical basics of microscopy focussing on the image capture and analysis (camera and chip technology, live cell imaging, analysis software). The characteristics of different sensor types were demonstrated by ANDOR and TCD staff in the in-house microscopy facilities and on image analysis workstations.

Outreach [top]
'The Complex Life of Sugars' is a comprehensive carbohydrate exhibition with a high degree of 'hands-on' activities for a broad range of audiences ranging from school children and A-level student over the general public to a scientific community. This stall showcases the science conducted within the group and links it to everyday life (biology, food and chemical as well as medical aspects). The ESR was involved in the organisation, demonstration and training of fellow presenters on three national occasions:

  1. MIB Open Day, 6th November 2015, UMAN, UK.
  2. Great British Bioscience Festival, BBSRC, 13-16th November 2014, London, UK.
  3. British Science Week 2015, 16-20th March 2015, UMAN, UK.

News

  • Discover Research Dublin night (September 25th 2015) An opportunity to share scientific research with the general public.
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  • Advanced Microscopy and Super Resolution (Andor) 07-09/05/15, Trinity College, Dublin: Lectures on the fundamentals of fluorescence imaging including a breakdown of different types of fluorescent probes including dyes, antibodies and fluorescent proteins were given.
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© 2017 TINTIN - A Marie Curie Initial Training Network Programme funded under Grant Agreement No. 608381