The Use of Recombinant Lectins for the Bioanalysis of Cell Surface Glycosylation

Session: 
S2.2 Protein N-glycosylation
Code: 
FL2.2.2
Location (hall): 
Mannose
Start/end time: 
Monday, July 1, 2019 - 15:20 to 15:25
Flávio
Ferreira

Flávio Ferreira1, Brendan O'Connor1, Dermot Walls1

1Dublin City University, Dublin, Ireland

Biological glycosylation is the process which adds specific sugars to other sugars, proteins and lipids [1]. Protein glycosylation is one of the most important post-translational modifications, which occurs in more than half of all proteins present in the human body. Abnormal glycosylation has been demonstrated to be linked to many different diseases due to alterations associated with protein folding and biological function [2]. Therefore, glycosylation is absolutely essential for the correct structure, function and stability of important proteins [3].

Surface glycosylation patterns play a key role in the modulation of the immune responses which are mediated by carbohydrate-binding proteins called Lectins [4]. Such biomolecules are typically highly selective for specific glycan structures, making them extremely useful for glycan variation investigation [5].

A rapid and accurate bioanalytical method to detect early unhealthy cell signs during a bioprocess is a current issue facing the industry. It is widely known that as cells become stressed or diseased the earliest changes that occur are in cell surface glycosylation.  

CHO cells are the host cell of choice of the rapidly emerging biopharmaceutical industry for the production of glycoprotein therapeutics. Hence, this research work investigated the interaction between recombinant lectin probes with glycoconjugates on the surface of the CHO-K1 cells and on secreted biomolecules. With the aid of computational biology techniques, statistically significant differences were found on cell surface glycoprofile in response to variation in temperature, CO2 and nutrient depletion levels of the cell culture process.   

References: 
  1. Marth, J. D.; Grewal, P. K. Mammalian glycosylation in immunity. Nature reviews. Immunology. 2008, 8, 874–887. 
  2. Christiansen, M. N.; Chik, J.; Lee, L.; Anugraham, M.; Abrahams, J. L.; Packer, N. H. Cell surface protein glycosylation in cancer. Proteomics. 2014, 14, 525–546. 
  3. Lepenies, B.; Seeberger, P. H. Simply better glycoproteins. Nature Biotechnology. 2014, 32, 443–445.
  4. Veiseh, M.; Kwon, D. H.; Borowsky, A. D.; Tolg, C.; Leong, H. S.; Lewis, J. D.; Turley, E. A.; Bissell, M. J. Cellular heterogeneity profiling by hyaluronan probes reveals an invasive but slow-growing breast tumor subset. Proc. Nat. Acad. Sci. U.S.A.[Online] 2014, 111, e1731–e1739 https://www.pnas.org/content/111/17/E1731 (accessed Feb 21, 2019).
  5. Ohtsubo, K.; Marth, J.D. Glycosylation in Cellular Mechanisms of Health and Disease. Cell. 2006 126, 855–867.

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