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.

1. Yang, Q.; Chen, H.; Ou, C.; Zheng, Z.; Zhang, X.; Liu, Y.; Zong, G.; Wang, L.X. Evaluation of Two Chemoenzymatic Glycan Remodeling Approaches to GenerateSite-Specific Antibody-Drug Conjugates. Antibodies(Basel) 2023, 12 (4).

2. Donahue, T. C.; Ou, C.; Yang, Q.; Flinko, R.; Zhang, X.; Zong, G.; Lewis, G.K.; Wang, L. X. Synthetic Site-Specific Antibody-Ligand Conjugates PromoteAsialoglycoprotein Receptor-Mediated Degradation of Extracellular Human PCSK9. ACS Chem Biol 2023, 18 (7), 1611-1623.

3. Zhang, X.; Ou, C.; Liu, H.; Wang, L. X. Synthesis and Evaluation of ThreeAzide-Modified Disaccharide Oxazolines as Enzyme Substrates for Single-Step FcGlycan-Mediated Antibody-Drug Conjugation. BioconjugChem 2022, 33 (6), 1179-1191.

4. Zhang, X.; Liu, H.; He, J.; Ou, C.; Donahue, T. C.; Muthana, M. M.; Su, L.;Wang, L. X. Site-Specific Chemoenzymatic Conjugation of High-Affinity M6PGlycan Ligands to Antibodies for Targeted Protein Degradation. ACS Chem Biol 2022, 17 (11), 3013-3023.

5.  Ou, C.; Prabhu, S. K.; Zhang, X.; Zong, G.; Yang, Q.; Wang, L. X. SyntheticAntibody-Rhamnose Cluster Conjugates Show Potent Complement-Dependent CellKilling by Recruiting Natural Antibodies. Chemistry(Weinheim an der Bergstrasse, Germany) 2022,28 (16), e202200146.

6. Zhang, X.; Ou, C.; Liu, H.; Prabhu, S. K.; Li, C.; Yang, Q.; Wang, L. X.General and Robust Chemoenzymatic Method for Glycan-Mediated Site-SpecificLabeling and Conjugation of Antibodies: Facile Synthesis of HomogeneousAntibody-Drug Conjugates. ACS Chem Biol 2021, 16 (11), 2502-2514.

7. Zhang, X.; Liu, H.; Meena, N.; Li, C.; Zong, G.; Raben, N.; Puertollano, R.;Wang, L. X. Chemoenzymatic glycan-selective remodeling of a therapeuticlysosomal enzyme with high-affinity M6P-glycan ligands. Enzyme substratespecificity is the name of the game. ChemSci 2021, 12 (37), 12451-12462.

8. Yang, Q.; Zhu, S.; Zhang, R.; Loke Chun, M.; Wang, L.-X.; An, Y.; Cipollo John,F. Glycan Remodeling of Human Erythropoietin (EPO) Through Combined MammalianCell Engineering and Chemoenzymatic Transglycosylation. ACS Chem Biol 2017, 12 (6), 1665-1673.

9. Yamaguchi, T.; Amin Mohammed, N.; Wang, L.-X.; Toonstra, C. ChemoenzymaticSynthesis and Receptor Binding of Mannose-6-Phosphate (M6P)-ContainingGlycoprotein Ligands Reveal Unusual Structural Requirements for M6P ReceptorRecognition. J Am Chem Soc 2016, 138 (38), 12472-12485.  

10. Li, T.; Tong, X.; Yang, Q.; Giddens, J. P.; Wang, L.-X. Glycosynthase Mutantsof Endoglycosidase S2 Show Potent Transglycosylation Activity and RemarkablyRelaxed Substrate Specificity for Antibody Glycosylation Remodeling. Journal of Biological Chemistry 2016, 291 (32), 16508-16518.

11. Giddens, J. P.; Lomino, J. V.; Amin, M. N.; Wang, L.-X. Endo-F3 GlycosynthaseMutants Enable Chemoenzymatic Synthesis of Core-fucosylated TriantennaryComplex Type Glycopeptides and Glycoproteins. Journal of Biological Chemistry 2016, 291 (17),9356-9370.

12. 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 glycanspecificity of HIV-neutralizing antibodies. NatureChemical Biology 2013, 9 (8), 521-526.

13. Huang, W.; Giddens, J.; Fan, S.-Q.; Toonstra, C.; Wang,L.-X. Chemoenzymatic Glycoengineering of Intact IgG Antibodies for Gain ofFunctions. Journal of the AmericanChemical Society 2012, 134 (29), 12308-12318.