‘I have umpteen thousand samples to analyse. How can I get reliable, detailed, quantitative maps of the complex glycosylation for each of them without spending a great deal of time and vast sums of money?’

 This talk gives an overview of LongBow - an integrated glycomics platform being developed to address this problem -  which is becoming increasingly important in two areas of modern healthcare:

a.  Biopharmaceutical  realisation .  Glycomics  studies are  essential  throughout  the    development  and biomanufacturing  of  most  glycoprotein  therapeutics because  changes  in  the  glycosylation  can  greatly influence the safety and efficacy of the drugs in patients.

b.  Clinical  diagnostics .  Glycans  play  key  roles  in  many critical biological processes and changes in glycosylation patterns  of  serum  glycoproteins  are  potential  early-warning biomarkers for a number of chronic diseases. These  include  cancers  and  serious  inflammatory conditions such as inflammatory bowel disease (IBD).In  both  cases,  we  are  interested  in  monitoring  the (often  subtle)  changes  in  glycosylation  patterns  that correlate  either  with  altered  clinical  performance  (for the  drugs)  or  disease  progress  in  the  patient.  However, determining glycosylation patterns is still very challenging because  of  the  complexity  and  heterogeneity  of  glycan populations.  The  complexity  arises  from  the  diversity  in glycan monomer types and the ways they can be assembled with  branched  topologies.  Regarding  heterogeneity,  both biopharmaceuticals and biological samples may be bear tens to hundreds of different types of glycans. For full glycomics studies we need to identify the complete structure of each glycan and determine its relative or absolute abundance to a high level of accuracy. Currently, there is no single analytical platform  that  can  do  this  reliably,  quickly  and  affordably. LongBow is being designed as a first step towards achieving this and implements concepts developed in the HighGlycan programme.  This  is  a  five  year,  €6m,  EU-FP7  funded collaborative  research  project  involving  nine  glycoscience organisations across Europe.The main topics covered in this overview will be:

1.  Overall  design  of  LongBow,  how  the  components  fit together, flexibility for chemistry and instrumentation

2.  Types  of  samples:  biopharmaceuticals,  plasma,  serum glycoproteins, cells and tissues

3.   Instrumentation  -  robotics  and  orthogonal  analytics (UHPLC, MS, LC/MS and xCE)

2.  Sample  processing  workflows  for  glycopeptides, N-glycans,  O-glycans,  sialic  acids  and  neutral monosaccharides

3.   Integration of modules, performance and validation

4.  VTag TM  system for glycopeptide analysis by UHPLC and MS

5.   Procainamide system for glycan analysis by UHPLC and MS

6.  Applications:  How  LongBow  is  being  used  at  Ludger for  biopharmaceutical  glycoprofiling  to  support  drug comparability  studies  and  regulatory  submissions  and for medical glycomics for stratification of patients with chronic inflammatory conditions

7.  Transfer of LongBow technology for glycomics studies in partner labs

8.  Advantages  and  limitations  of  LongBow,  future improvements.