Training

Our two-week training courses are run in July and August each summer.  If you would like to be considered for summer 2012 please email Anne Dell at a.dell@imperial.ac.uk attaching a CV and a brief account of your reasons for applying.

We do not charge any fees for the training courses but we are unable to fund travel or accommodation.  Find out about inexpensive short-term College accommodation here

Training

What is Glycobiology?

Glycobiology encompasses the disciplines of glycomics and glycoproteomics. These are the study of carbohydrates (glycans, sugars, saccharides) attached to proteins. More than half of all proteins are predicted to be glycoproteins. The role of glycans impinges on many in vivo events, including:

• Cell adhesion e.g. lymphocyte rolling
• Immunological events
• Hepatic Hormone Clearance
• Neural development/function

Accordingly full structural definition of glycoproteins is a desirable objective in many fields of biology and medicine. Establishment of the complete glycan repertoires form an important part of our understanding of structure-function relationship. It is hoped that such understanding will allow us to modulate fundamental biological systems for beneficial effects.  In many circumstances, owing to low sample quantities and structural heterogeneity, mass spectrometry-based glycomics/ glycoproteomics is the method of choice for structural analysis.

Frequently Asked Questions

When do courses run?
July and August but training may be arranged at other times during the year if resources are available

How long is the course?
Two or three weeks. 

How can I find accommodation?
We ourselves do not have the resources to advise on/arrange accommodation, but the college does provide advice on accommodation for visitors to the college. Follow this link: http://www3.imperial.ac.uk/accommodation/visitingimperial

What are the course fees?
Presently, it is FREE to enrol on the GlycoTric course. All we ask is for you to fill out our in-course questionnaire at the end of the course as we are constantly seeking to improve the course

Do I need previous knowledge?
No, we cater for the complete novice as well as those with more experience.

Am I guaranteed a place on the course?
No, because resources are limited. Therefore we request a C.V. and your reasons for wanting our training.

What are the main course objectives?
Hands-on-training in glycoprotein and glycomics with an emphasis on sample preparation, including:
• Cell extractions
• Tissue extractions
• Analysis of a standard glycoprotein
• Carboxymethylation
• Tryptic digestion
• N- and O-glycan release
• Permethylation
• Sep-Pak purication of peptides/glycopeptides and permethylated samples
• N- and O-glycan MS profiling
• MS/MS of  N- and O-glycans
• Data interpretation of MS data
• Data interpretation of MS/MS data
• An outline of web-based glycomics resources
• TMS sugar analysis (if requested)
• Data interpretation of nano-LC-MS/MS data (if requested)

How long does it take to prepare a glycoprotein for glycomics analysis?
One week to release and analyse O-glycans from glycoproteins
Two weeks to release and analyse N-glycans from glycoproteins
Two and a half weeks to release N-and O-glycans from the same glycprotein

What are the benefits of NaOH permethylation of carbohydrates?
Increases mass spectrometric sensitivity by an order of magnitude.
Esterifies sialic acid carboxylic acid groups which stabilise these labile groups. This gives a better representation of sialic acid content which is typically under represented in other methodologies.
Directs fragmentation of carbohydrates in MS/MS experiments. This aids interpretation.

What about sulphated carbohydrates?
Sulphated species are lost in the workup of the standard NaOH permethylation procedure. We can advice on analysis of sulphated samples but hands-on training is not provided at present.

Will I get a certificate?
Yes, all attendees who satisfactorily complete the course will receive a GlycoTric certificate.

Hands-on training may involve several hazards. Will I get appropriate advice?
Laboratories are potentially hazardous places. We ask you to read and follow our rules of good laboratory practice before entering the laboratory.  COSHH documentation will be available for all chemicals that are used. This will be explained on arrival.
 
WE RESERVE THE RIGHT TO SUSPEND TRAINING IF A PARTICIPANT DOES NOT COMPLY WITH GOOD LABORATORY PRACTICE

About our Training

The integrated nature of our strategies allows us to address some of the most  taxing glycomic/ glycoproteomic structural problems. We have incorporated elements of these strategies into the programme making it suitable for experienced participants aiming to optimise their skills as well as complete novices who are new to this exciting area of research. We believe that hands-on training is the most effective means of training and it forms the core of the programme. Most trainees enrol on a two week programme and choose to analyse a selection of samples that may include any of the following:

• Permethylation of a sugar standard and MALDI-TOF/TOF MS/MS
• Immunoglobin A - human (N- and O- glycan analysis)      
• Immunoglobin G - human (N- glycan analysis)
• Fetuin - Fetal calf (N- and O- glycans)
• Tissue samples (N- and O- glycans)
• Cell samples (N- and O- glycans)

We also actively encourage people to bring their own samples. In the summer session of 2007 the samples that were analysed were quite diverse ranging from mucins from an ovarian cyst to glycoprotein extract from a sea urchin spines. Modified cell cells such as the following are also suitable for analysis:

• Leukocytes (neutrophils, B- and T- lymphocyrtes, macrophages etc)
• Modified CHO cell lines
• Miscellaneous cell lines e.g. HEK293 cells
• Modified bacterial glycoproteins from E.coli

What Our Courses Offer

Hands-on training for mass spectrometric preparation for glycoprotein, tissue or own  samples.  Preparation of N-glycans and O- glycans:

• Hands-on training for derivatisation and sample loading
• Demonstration of MALDI-TOF MS  and MALDI-TOF/TOF MS/MS 
• MALDI-TOF MS Data Interpretation
• MALDI-TOF/TOF MS/MS Data Interpretation
• Demonstrations of glycoproteomic technology

We are experts in the challenges of micro-scale sample handling, a necessary requirement for successful mass spectrometric analysis. We are also aware of the apparent complexities of data interpretation at both novice and expert levels. We understand this, as this is what we do every day, and have done for many years.  To this end, we provide an informative training programme that we hope is both helpful and enjoyable.