GlycoT Therapeutics

Technology

 

The key technology of GlycoT (developed by Prof. Wang’s group in University of Maryland) permits chemoenzymatic glycoengineering of therapeutic antibodies and other glycoprotein products to provide structurally well-defined, homogeneous glycoforms with improved or gain of functionalities. This technology development has resulted in over a dozen of approved US and international patents. The IPs cover the novel enzymes/mutants, the process, and the products. As demonstrated for the glycosylation remodeling of antibodies, this technology consists of two key steps: deglycosylation of the antibody by an endoglycosidase to remove the heterogeneous Fc glycans and subsequent attachment of a desired native or modified N-glycan to the deglycosylated antibody by a novel glycosynthase to reconstitute the antibody. This technology opens a new avenue to accessing a wide range of homogeneous antibody glycoforms , including Fc-glycan specific antibody drug conjugates, that would be otherwise difficult to obtain by other chemical or biological approaches.

Most recently, Wang lab has developed another one-pot transglycosylation technology that can introduce functionalized synthetic disaccharides to antibody with just one Endo-S2 enzyme in a single step reaction, as shown below. Drugs or other functional molecules can be introduced subsequently with mild click reactions. The technology has been exclusively licensed by GlycoT.

Selected Reference

  1. Zhang X, Ou C, Liu H, Prabhu SK, Li C, Yang Q, Wang LX. General and Robust Chemoenzymatic Method for Glycan-Mediated Site-Specific Labeling and Conjugation of Antibodies: Facile Synthesis of Homogeneous Antibody-Drug Conjugates. ACS Chem Biol. 2021;16(11):2502-2514.
  2. Ou C, Li C, Zhang R, Yang Q, Zong G, Dai Y, Francis RL, Bournazos S, Ravetch JV, Wang LX. One-Pot Conversion of Free Sialoglycans to Functionalized Glycan Oxazolines and Efficient Synthesis of Homogeneous Antibody-Drug Conjugates through Site-Specific Chemoenzymatic Glycan Remodeling. Bioconjug Chem. 2021;32(8):1888-1897.
  3. Yang Q, An Y, Zhu S, Zhang R, Loke CM, Cipollo JF, Wang LX, “Glycan Remodeling of Human Erythropoietin (EPO) Through Combined Mammalian Cell Engineering and Chemoenzymatic Transglycosylation”, ACS Chem. Biol., 12, 1665-1673 (2017).
  4. Li, T., DiLillo, D., Bournazos, S., Giddens, J. P., Ravetch, J. V., Wang, L. X., “Modulating IgG effector functions by Fc glycan engineering”, Proc. Natl. Acad. Sci. USA, 114, 3485-3490 (2017).
  5. Yamaguchi, T., Amin, M.N., Toonstra, C., Wang, L.X., “Chemoenzymatic synthesis and receptor binding of mannose-6-phosphate (M6P)-containing glycoprotein ligands reveal unusual structural requirements for M6P receptor recognition”,  J. Am. Chem. Soc., 138, 12472-85 (2016).
  6. Li, T., Tong, X., Yang, Q., Giddens, J.P., Wang, L.X., “Glycosynthase mutants of endoglycosidase S2 show potent transglycosylation activity and remarkably relaxed substrate specificity for antibody glycosylation remodeling”, J Biol. Chem., 291, 16508-16518 (2016)
  7. Giddens, J.P., Lomino, J.V., Amin, M.N., Wang, L.X., “Endo-F3 glycosynthase mutants enable chemoenzymatic synthesis of core fucosylated tri-antennary complex-type glycopeptides and glycoproteins”, J. Biol. Chem., 291, 11064-11071 (2016).
  8. Amin, M.N., McLellan, J.S., Huang, W., Orwenyo, J., Burton, D.R., Koff, W.C., Kwong, P.D., Wang, L.X., “Synthetic glycopeptides reveal the glycan specificity of HIV-neutralizing antibodies”, Nature Chem. Biol., 9, 521-526 (2013).
  9. Huang, W., Giddens, J., Fan, S.Q., Toonstra, C., Wang, L.X., “Chemoenzymatic glycoengineering of intact IgG antibodies for gain of functions”, JAm. Chem. Soc., 134, 12308−12318 (2012).