Broad-Spectrum Legionaminic Acid-Specific Antibodies in Pooled Human IgGs Revealed by Glycan Microarrays with Chemoenzymatically Synthesized Nonulosonosides
Abstract
:1. Introduction
2. Results and Discussion
2.1. Chemoenzymatic Synthesis of Leg5,7Ac2-Glycosides
2.2. Glycan Microarray Binding Studies Reveal a Broad Spectrum of Human IgG Antibodies against Leg5,7Ac2-Glycosides
3. Materials and Methods
3.1. Materials and Instruments
3.2. General Procedures for One-Pot Three-Enzyme (OP3E) Preparative-Scale Synthesis of Leg5,7diN3α2–6-Linked Nonulosonosides (12–18)
3.3. General Procedures for One-Pot Three-Enzyme (OP3E) Preparative-Scale Synthesis of Leg5,7diN3α2–3-Linked Nonulosonosides (19–24)
3.4. General Procedures for Synthesizing Leg5,7Ac2-Glycosides (25–37) by Converting the Azido Groups in Leg5,7diN3-Glycosides (12–24) to NHAc Groups
3.5. General Procedures for Converting Cbz-Tagged Glycans to Glycosyl Propylamines
3.6. Glycan Microarray Fabrication
3.7. Glycan Microarray Binding Assays
3.8. Array Slide Processing
3.9. Statistics
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Acceptors (5–11) | OP3E Products Leg5,7diN3-glycosides (12–24) | Leg5,7Ac2-glycosides (25–37) | ||
12–18 (70–99%) | 19–24 (71–96%) | 25–31 (65–90%) | 32–37 (67–75%) | |
GalNAcαProNHCbz (5) R = αProNHCbz, R1 = NHAc | 12 (99%) | - | 25 (65%) | - |
R = β4GlcβProNHCbz, R1 = OH | 13 (75%) | 19 (74%) | 26 (78%) | 32 (68%) |
R = β4GlcNAcβProNHCbz, R1 = OH | 14 (70%) | 20 (71%) | 27 (68%) | 33 (67%) |
R = β3GalNAcβProNHCbz, R1 = OH | 15 (93%) | 21 (96%) | 28 (90%) | 34 (67%) |
R = β3GalNAcαProNHCbz, R1 = OH | 16 (77%) | 22 (77%) | 29 (68%) | 35 (69%) |
R = β3GlcNAcβProNHCbz, R1 = OH | 17 (81%) | 23 (80%) | 30 (78%) | 36 (75%) |
R = β3GlcNAcαProNHCbz, R1 = OH | 18 (99%) | 24 (96%) | 31 (71%) | 37 (72%) |
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Kooner, A.S.; Yu, H.; Leviatan Ben-Arye, S.; Padler-Karavani, V.; Chen, X. Broad-Spectrum Legionaminic Acid-Specific Antibodies in Pooled Human IgGs Revealed by Glycan Microarrays with Chemoenzymatically Synthesized Nonulosonosides. Molecules 2024, 29, 3980. https://doi.org/10.3390/molecules29163980
Kooner AS, Yu H, Leviatan Ben-Arye S, Padler-Karavani V, Chen X. Broad-Spectrum Legionaminic Acid-Specific Antibodies in Pooled Human IgGs Revealed by Glycan Microarrays with Chemoenzymatically Synthesized Nonulosonosides. Molecules. 2024; 29(16):3980. https://doi.org/10.3390/molecules29163980
Chicago/Turabian StyleKooner, Anoopjit Singh, Hai Yu, Shani Leviatan Ben-Arye, Vered Padler-Karavani, and Xi Chen. 2024. "Broad-Spectrum Legionaminic Acid-Specific Antibodies in Pooled Human IgGs Revealed by Glycan Microarrays with Chemoenzymatically Synthesized Nonulosonosides" Molecules 29, no. 16: 3980. https://doi.org/10.3390/molecules29163980