Professor Chi-Huey WONG's Laboratory

Group Focus

Research in Professor Wong's lab. encompasses a broad spectrum of bioorganic and synthetic chemistry, aiming to understand carbohydrates in biology and develop new therapeutic strategies and devices. Development of novel molecular structures to investigate how they interact with cells and learn more about the mechanism and function of those molecules and disease progression. Programmable one-pot reactions are being developed for the synthesis of complex oligosaccharides and glycoarrays and complement new chemo-enzymatic strategies for the synthesis of glycoproteins, vaccines and other biologically active molecules to tackle major problems in biology and medicine, especially those associated with cancer and infectious diseases.

About Professor Chi-Huey WONG

[CV][Wikipedia]

Education

Academic Appointments

Awards

Research Interests

Click [Google Scholar] to see our all publications.

Chemistry and Chemical Biology of Biological Glycosylation

Research in the Wong lab encompasses a broad spectrum of bioorganic, synthetic, and AI-assisted chemistry, aiming to understand the role of carbohydrates in biology and develop new therapeutic strategies and devices. Development of novel molecular structures to investigate how they interact with cells and learn more about the mechanism and function of those molecules and disease progression is of current interest. Programmable one-pot reactions are being developed for the synthesis of complex oligosaccharides and glycan microarrays and complement new chemo-enzymatic strategies for the synthesis of complex carbohydrates, glycoproteins, vaccines, enzyme inhibitors and other biologically active molecules to tackle major problems in biology and medicine, especially those associated with cancer and infectious diseases.

Glycan Synthesis

The access to the structure-defined glycan samples in a pure (homogeneous) form is instrumental to the study of glycosylation and the development of therapeutic agents targeting the glycosylation processes associated with cancer, as well as glycosylation patterns associated with bacterial and viral infections. Currently, chemical and enzymatic syntheses are the only effective methods for producing homogenous glycans. To simplify glycan synthesis, we developed a programmable one-pot method for chemical synthesis of oligosaccharides and an enzymatic method for the synthesis of oligosaccharides with regeneration of sugar nucleotide cofactor. We use the synthetic glycans in biological assays, glycan microarrays and for the development of therapeutic vaccines, and glycoengineered proteins.

• Cheng, C. W.; Zhou, Y.; Pan, W. H.; Dey, S.; Wu, C. Y.; Hsu, W. L.; Wong, C. H. "Hierarchical and programmable one-pot synthesis of oligosaccharides" Nat. Commun. 2018, 9, 5202.
• Dey, S.; Lo, H. J.; Wong, C. H. "An Efficient Modular One-Pot Synthesis of Heparin-Based Anticoagulant Idraparinux" J. Am. Chem. Soc. 2019, 141, 10309-10314.
• Huang, L.-Y.; Huang, S.-H.; Chang, Y.-C.; Cheng, W.-C.; Cheng, T.-J.; Wong, C.-H. "Enzymatic synthesis of lipid II and analogues" Angew. Chem. Int. Ed. Engl. 2014, 53, 8060-8065.
• Shivatare, S. S.; Chang, S.-H.; Tsai, T.-I.; Tseng, S.-Y.; Shivatare, V.-S.; Lin, Y.-S.; Cheng, Y.-Y.; Ren, C.-T.; Lee, C.-C.; Pawar, S.; Tsai, C.-S.; Shih, H.-W.; Zeng, Y.-F.; Liang, C.-H.; Kwong, P.-D.; Burton, D.-R.; Wu, C.-Y.; Wong, C.-H. "Modular synthesis of N-glycans and arrays for the hetero-ligand binding analysis of HIV antibodies" Nat. Chem. 2016, 8, 338-346.
• C.-W. Chang, M.-H. Lin, C.-K. Chan, K.-Y. Su, C.-H. Wu, W.-C. Lo, S. Lam, Y.-T. Cheng, P.-H. Liao, C.-H. Wong, C.-C. Wang, “Automated quantification of hydroxyl reactivities: prediction of glycosylation reactions” Angew. Chem. Int. Ed. 2021, 60, 12413-12423.
• S. S. Shivatare, V. S. Shivatare and C.-H. Wong, “Glycoconjugates: synthesis, functional studies and therapeutic developments” Chem. Rev. 2022, 122, 15603-15671.

Homogenous and Specific Glycoform-enriched Glycoproteins and Antibodies

Despite recent advances in genetic engineering of glycosylation pathways, the recombinant glycoproteins are generally obtained as mixtures of glycoforms making them unsuitable for functional studies. The Wong group was the first to realize enzymatic glycan remodeling to produce homogeneous glycoproteins demonstrating functional and structural differences between different glycoforms. The monoclonal antibody, immunoglobulin G, is one of the prominent glycoproteins, whose effector functions are regulated by N-glycosylation of Fc domain. In our group, we study the glycosylation of therapeutic monoclonal antibodies to improve their Fc-receptor binding and potency and develop enzymatic and cell-based glycoengineering methods to produce homogeneous or specific-glycoform-enriched antibodies with improved effector functions, including antibody-dependent cellular cytotoxicity (ADCC) and phagocytosis (ADCP) as well as antibody-dependent vaccinal effect (ADVE).

• Witte, K.; Sears, P.; Martin, R.; Wong, C.-H. "Enzymatic glycoprotein synthesis: preparation of ribonuclease glycoforms via enzymatic glycopeptide condensation and glycosylation" J. Am. Chem. Soc. 1997, 119, 2114-2118.
• Culyba, E. K.; Price, J. L.; Hanson, S. R.; Dhar, A.; Wong, C.-H.; Gruebele, M.; Powers, E. T.; Kelly, J. W. "Protein native state stabilization by placing aromatic side chains in N-glycosylated reverse turns" Science 2011, 331, 571-575.
• Lin, C.-W.; Tsai, M.-H.; Li, S.-T.; Tsai, T.-I.; Chu, K.-C.; Liu, Y.-C.; Lai, M.-Y.; Wu, C.-Y.; Tseng, Y.-C.; Shivatare, S.-S.; Wang, C.-H.; Chao, P.; Wang, S.-Y.; Shih, H.-W.; Zeng, Y.-F.; You, T.-H.; Liao, J.-Y.; Tu, Y.-C.; Lin, Y.-S.; Chuang, H.-Y.; Chen, C.-L.; Tsai, C.-S.; Huang, C.-C.; Lin, N.-H.; Ma, C.; Wu, C.-Y.; Wong, C.-H. "A common glycan structure on immunoglobulin G for enhancement of effector functions." Proc. Natl. Acad. Sci. U. S. A. 2015, 112, 10611-10616.
• Lo, H. J.; Krasnova, L.; Dey, S.; Cheng, T.; Liu, H.; Tsai, T. I.; Wu, K. B.; Wu, C. Y.; Wong, C. H. "Synthesis of Sialidase-Resistant Oligosaccharide and Antibody Glycoform Containing alpha2,6-Linked 3F(ax)-Neu5Ac" J. Am. Chem. Soc. 2019, 141, 6484-6488.
• V. S. Shivatare, P.-K Chuang, T.-H. Tseng, Y.-F. Zeng, H.-W. Huang, G. Veeranjaneyulu, H.-C. Wu, C.-H. Wong, “Study on antibody Fc-glycosylation for optimal effector functions” ChemComm. 2023, 59, 5555-5558.
• H. W. Huang, Y. F. Zeng, V. S. Shivatare, T. H. Tseng, and C. H. Wong, “Cell-based glycoengineering for production of homogeneous and specific glycoform-enriched antibodies with improved effector functions” Proc. Natl. Acad. Sci. USA. 2025, Doi: 10.1073/pnas.2423853122.

Glycosylation Probes

The glycosylation probes are monosaccharides modified with a functional group (e.g., azide or alkyne), which could be labeled with a reporter tag using bioorthogonal chemistry. Metabolic incorporation of glycosylation probes allows monitoring of glycosylation processes in the cell and in vivo. By comparing the differences in labeling between healthy cells and cancer cells, we can identify the cancer-specific glycan biomarkers and develop new diagnostic tools, as well as discover new targets for the development of therapeutic agents. In our group, we are focusing on the alkyne-bearing probes with improved sensitivity and cell permeability, as well better biorthogonal labeling suitable for real-time imaging.

• Sawa, M.; Hsu, T. L.; Itoh, T.; Sugiyama, M.; Hanson, S. R.; Vogt, P. K.; Wong, C. H. "Glycoproteomic probes for fluorescent imaging of fucosylated glycans in vivo" Proc. Natl. Acad. Sci. U. S. A. 2006, 103, 12371-12376.
• Kizuka, Y.; Funayama, S.; Shogomori, H.; Nakano, M.; Nakajima, K.; Oka, R.; Kitazume, S.; Yamaguchi, Y.; Sano, M.; Korekane, H.; Hsu, T. L.; Lee, H. Y.; Wong, C. H.; Taniguchi, N. "High-Sensitivity and Low-Toxicity Fucose Probe for Glycan Imaging and Biomarker Discovery" Cell Chem. Biol. 2016, 23, 782-792.
• Tsai, C.-S.; Yen, H.-Y.; Lin, M.-I.; Tsai, T.-I.; Wang, S.-Y.; Huang, W.-I.; Hsu, T.-L.; Cheng, Y.-S. E.; Fang, J.-M.; Wong, C.-H. "Cell-permeable probe for identification and imaging of sialidases" Proc. Natl. Acad. Sci. U. S. A. 2013, 110, 2466-2471.
• Shie, J.-J.; Liu, Y.-C.; Lee, Y.-M.; Lim, C.; Fang, J.-M.; Wong, C.-H. "An azido-BODIPY probe for glycosylation: initiation of strong fluorescence upon triazole formation" J. Am. Chem. Soc. 2014, 136, 9953-9961.
• S.-W. Wang, Y.-A. Ko, C.-Y. Chen, K.-S. Liao, Y.-H. Chang, H.-Y. Lee, Y.-H. Yu, Y.-H. Lih, Y.-Y. Cheng, H.-H. Lin, T.-L. Hsu, C.-Y. Wu, K.-I Lin, C.-H. Wong, “Mechanism of antigen presentation and specificity of antibody cross-reactivity elicited by an oligosaccharide-conjugate cancer vaccine” J. Am. Chem. Soc. 2023, 145, 9840-9849.

Glyco-enzyme Inhibitors

The inhibitors of glycan-processing enzymes could be used as probes to study glycosylation processes with the ultimate goal to develop new therapeutics. The enzymatic targets pursued in our group are comprised of influenza neuraminidase, bacterial transglycosylase, and mammalian b3GalT5 which catalyzes the galactosylation of Gb4 to generate the cancer-associated Gb5 (SSEA3). In our approach, we use both the substrate-based design of inhibitors and structure optimization of hits from high throughput screening.

• Kizuka, Y.; Nakano, M.; Yamaguchi, Y.; Nakajima, K.; Oka, R.; Sato, K.; Ren, C. T.; Hsu, T. L.; Wong, C. H.; Taniguchi, N. "An Alkynyl-Fucose Halts Hepatoma Cell Migration and Invasion by Inhibiting GDP-Fucose-Synthesizing Enzyme FX, TSTA3" Cell Chem. Biol. 2017, 24, 1467-1478.
• Wang, P. C.; Fang, J. M.; Tsai, K. C.; Wang, S. Y.; Huang, W. I.; Tseng, Y. C.; Cheng, Y. S.; Cheng, T. J.; Wong, C. H. "Peramivir Phosphonate Derivatives as Influenza Neuraminidase Inhibitors" J. Med. Chem. 2016, 59, 5297-5310.
• Wu, W. S.; Cheng, W. C.; Cheng, T. R.; Wong, C. H. "Affinity-Based Screen for Inhibitors of Bacterial Transglycosylase" J. Am. Chem. Soc. 2018, 140, 2752-2755.
• Mitchell, M. L.; Tian, F.; Lee, L. V.; Wong, C. H. "Synthesis and evaluation of transition-state analogue inhibitors of alpha-1,3-fucosyltransferase" Angew. Chem. Int. Ed. Engl. 2002, 41, 3041-3044.
• J M Lo, CC Kung, TJ Cheng, CH Wong, C Ma, “Structure-based mechanism and specificity of human galactosyltransferase β3GalT5” J. Am. Chem. Soc. 2025, Doi:10.1021/jacs.4c11724.

Glycan Microarrays

Glycan microarrays are useful tools, in which the glycan arrangement mimics that of the cell surface glycans involved in the multivalent glycan-protein interactions. The Wong group pioneered the use of glycan microarray for the profiling of glycan-protein interactions and quantification of these interactions for biochemical studies and diagnostics. To improve the sensitivity of detection, we designed aluminum oxide-coated glass slides for immobilization of glycan samples through phosphonate chemistry. In our laboratory, glycan arrays were used to identify cancer-specific glycan markers and unusual hybrid-type N-glycan with high binding avidity to anti-HIV broadly neutralizing antibodies. The newly identified cancer-specific glycan biomarkers and glycan binders of anti-HIV antibodies are being investigated as haptens for the development of carbohydrate-based therapeutic vaccines.

• Fazio, F.; Bryan, M. C.; Blixt, O.; Paulson, J. C.; Wong, C. H. "Synthesis of sugar arrays in microtiter plate" J. Am. Chem. Soc. 2002, 124, 14397-14402.
• Wang, C. C.; Huang, Y. L.; Ren, C. T.; Lin, C. W.; Hung, J. T.; Yu, J. C.; Yu, A. L.; Wu, C. Y.; Wong, C. H. "Glycan microarray of Globo H and related structures for quantitative analysis of breast cancer" Proc. Natl. Acad. Sci. U. S. A. 2008, 105, 11661-11666.
• Chang, S. H.; Han, J. L.; Tseng, S. Y.; Lee, H. Y.; Lin, C. W.; Lin, Y. C.; Jeng, W. Y.; Wang, A. H.; Wu, C. Y.; Wong, C. H. "Glycan array on aluminum oxide-coated glass slides through phosphonate chemistry" J. Am. Chem. Soc. 2010, 132, 13371-13380.
• Shivatare, V. S.; Shivatare, S. S.; Lee, C.-D.; Liang, C.-H.; Liao, K.-S.; Cheng, Y.-Y.; Saidachary, G.; Wu, C.-Y.; Lin, N.-H.; Kwong, P. D.; Burton, D. R.; Wu, C.-Y.; Wong, C.-H. "Unprecedented role of hybrid N-glycans as ligands for HIV-1 broadly neutralizing antibodies" J. Am. Chem. Soc. 2018, 140, 5202-5210.

Low-sugar Universal Vaccines against Cancer and Viral Pathogens

Cancer progression is associated with the altered composition of glycolipids or glycoproteins. In one of our studies, we established that globo-series glycans (Globo-H, SSEA-3, and SSEA-4) are found on the surface of 15 different types of cancers, including the breast cancer and breast cancer stem cells. Since these glycans are specific to cancerous cells and can stimulate an immune response after conjugation to a carrier protein, they are being investigated as haptens for therapeutic vaccines against cancer. Some of the disadvantages of carbohydrate-based vaccines include reduced immunogenicity and inability to generate a long-lasting response. Overcoming these challenges, we improved the formulation of Globo-H vaccine with diphtheria toxoid cross-reactive material 197 as an immunogenic carrier protein, and α-Gal ceramide analog C34 as an adjuvant to activate the IgM to IgG switch. In addition to the development of broadly effective vaccines and antibodies for the treatment of various cancers, our group is also developing universal vaccines against influenza virus, SARS-CoV-2, HIV and mutants based on our understanding of their mutation and glycosylation effect on the immunogenicity of viral cell-surface proteins. We found that the highly conserved epitopes of influenza hemagglutinin and SARS-CoV-2 spike protein are shielded by host-made glycans and deletion of the glycan shields expose the conserved epitopes and elicit broadly protective immune responses against different variants, providing a new approach to development of universal vaccines.

• Danishefsky, S. J.; Shue, Y.-K.; Chang, M. N.; Wong, C.-H. "Development of Globo-H cancer vaccine" Acc. Chem. Res. 2015, 48, 643-652.
• Huang, Y.-L.; Hung, J.-T.; Cheung, S.-K.; Lee, H.-Y.; Chu, K.-C.; Li, S.-T.; Lin, Y.-C.; Ren, C.-T.; Cheng, T.-J.; Hsu, T.-L.; Yu, A.-L.; Wu, C.-Y.; Wong, C.-H. "Carbohydrate-based vaccines with a glycolipid adjuvant for breast cancer" Proc. Natl. Acad. Sci. U. S. A. 2013, 110, 2517-2522.
• Tseng, Y. C.; Wu, C. Y.; Liu, M. L.; Chen, T. H.; Chiang, W. L.; Yu, Y. H.; Jan, J. T.; Lin, K. I.; Wong, C. H.; Ma, C. "Egg-based influenza split virus vaccine with monoglycosylation induces cross-strain protection against influenza virus infections" Proc. Natl. Acad. Sci. U. S. A. 2019, 116, 4200-4205.
• Liao, H.-Y.; Krasnova, L.; Wong, C.-H. "Chimeric hemagglutinin vaccine elicits broadly protective CD4 and CD8 T cell responses against multiple influenza strains and subtypes" Proc. Natl. Acad. Sci. U. S. A. 2020, 117, 17757-63.
• H.-Y. Huang, H.-Y. Liao, X. Chen, C.-W. Cheng, S.-W. Wang, M. Shahed-Al-Mahmud, T.-H. Chen, J. M. Lo, Y.-M. Liu, H.-H. Ma, Y.-H. Chang, C.-Y. Tsai, P.-Y. Huang, S.-Y. Chang, T.-L. Chao, H.-C. Kao, Y.-M. Tsai, Y.-H. Chen, C.-Y. Chen, K.-C. Lee, C.-Y. Wu, J.-T. Jan, K.-I. Lin, T.-J. R. Cheng, C. Ma, and C.-H. Wong, “Vaccination with SARS-CoV-2 spike protein lacking glycan shields elicits enhanced protective responses in animal models" Sci. Transl. Med. 2022, 14, eabm0899.
• C.-W. Cheng, C.-Y. Wu, S.-W. Wang, J.-Y. Chen, C.-C. Kung, K.-S. Liao, C.-H. Wong, “Low-Sugar Universal mRNA Vaccine against Coronavirus Variants with Deletion of Glycosites in the S2 or Stem of SARS-CoV-2 Spike mRNA” Proc. Natl. Acad. Sci. U. S. A. 2023, 120, e2314392120.
• C.-Y. Wu, C.-W. Cheng, C.-C. Kung, K.-S. Liao, J.-T. Jan, C. Ma, C.-H. Wong, “Glycosite-deleted mRNA of SARS-CoV-2 Spike Protein as Broad-Spectrum Vaccine” Proc. Natl. Acad. Sci. U. S. A. 2022, 119, e2119995119.
• C.-Y. Chen, Y.-W. Lin, S.-W. Wang, Y.-C. Lin, Y.-Y. Cheng, C.-T. Ren, C.-H. Wong, and C.-Y. Wu, “Synthesis of Azido-Globo H Analogs for Immunogenicity Evaluation” ACS Cent. Sci. 2022, 8, 77-85.
• S.-W. Wang, Y.-A. Ko, C.-Y. Chen, K.-S. Liao, Y.-H. Chang, H.-Y. Lee, Y.-H. Yu, Y.-H. Lih, Y.-Y. Cheng, H.-H. Lin, T.-L. Hsu, C.-Y. Wu, K.-I. Lin, C.-H. Wong, “Mechanism of antigen presentation and specificity of antibody cross-reactivity elicited by an oligosaccharide-conjugate cancer vaccine” J. Am. Chem. Soc. 2023, 145, 9840-9849.
• J.-R. Chen, Y.-H. Yu, Y.-C. Tseng, W.-L. Chiang, M.-F. Chiang, Y.-A. Ko, Y.-K. Chiu, S.-H. Ma, C.-Y. Wu, J.-T. Jan, K.-I. Lin, C. Ma and C.-H. Wong, “Vaccination of monoglycosylated hemagglutinin induces cross-strain protection against influenza virus infections” Proc. Natl. Acad. Sci. U. S. A. 2014, 111, 2476-2481.

Group Members

Lab Manager

• Yu-Ya Kao

Postdoctoral Fellows

• Nai-Yu Chen
• Li-Chun Cheng
• Cinya Chung
• Srinivasa Rao Palla

Research Assistants

• Chi-Chuan Kung

Graduate Students

• Pony Chen
• Mien-Yao Peng
• Yang Huang

Contacts

Professor Chi-Huey WONG

Ms. Sylvia LIN

Professor Chi-Huey WONG's Laboratory, Genomics Research Center, Academia Sinica, Taiwan