Volume 2, Issue 1, Pages 114-124 (January 2017) Automated Glycan Assembly of Oligo-N-Acetyllactosamine and Keratan Sulfate Probes to Study Virus-Glycan Interactions  Heung Sik Hahm, Felix Broecker, Fumiko Kawasaki, Mario Mietzsch, Regine Heilbronn, Minoru Fukuda, Peter H. Seeberger  Chem  Volume 2, Issue 1, Pages 114-124 (January 2017) DOI: 10.1016/j.chempr.2016.12.004 Copyright © 2017 Elsevier Inc. Terms and Conditions

Chem 2017 2, 114-124DOI: (10.1016/j.chempr.2016.12.004) Copyright © 2017 Elsevier Inc. Terms and Conditions

Figure 1 Chemical Structures of LacNAc and KS Oligosaccharide Targets Chem 2017 2, 114-124DOI: (10.1016/j.chempr.2016.12.004) Copyright © 2017 Elsevier Inc. Terms and Conditions

Figure 2 Retrosynthetic Analysis of Linear, Branched, and Sulfated LacNAc and KS Oligosaccharides and the Complete Set of Building Blocks for AGA via the Reported Strategy with Three Orthogonal Temporary Protecting Groups Chem 2017 2, 114-124DOI: (10.1016/j.chempr.2016.12.004) Copyright © 2017 Elsevier Inc. Terms and Conditions

Figure 3 Binding Patterns of AAV Particles to 1–7 as Determined by Glycan Microarray and SPR Experiments (A) Exemplary microarray scan of fluorescence-labeled AAVrh10 incubated at 100 μg/mL. The spotting pattern of 1–7 and synthetic dermatan sulfate glycans53 S12 and S13 is indicated (see Figure S19 for further information). Each compound was spotted in triplicate. Binding of AAVrh10 to 6 is seen as three white spots, which represent fluorescence signals excited at 488 nm. (B) Binding signals of AAVrh10 to 6 as quantified by glycan microarray. Error bars represent mean + SEM of mean fluorescence intensity signals of six microarray spots. (C) Microarray-inferred binding signals of different AAV serotypes (100 μg/mL) and GS-I lectin (20 μg/mL) to the indicated glycans depicted in (B). The bars show mean plus standard deviation (S.D.) of three microarray spots. (D) SPR sensorgrams of non-labeled AAVrh10, which was passed at the indicated concentrations over sensor chip surfaces functionalized with either 6 (left) or 5 (right). The depicted sensorgrams are representative of two independent experiments with similar results. Chem 2017 2, 114-124DOI: (10.1016/j.chempr.2016.12.004) Copyright © 2017 Elsevier Inc. Terms and Conditions

Scheme 1 AGA of Linear and Branched LacNAc Oligosaccharides (A) The photolabile linker 8 and building blocks 9, 11, and 12. (B) Automated assembly. Reagents and conditions: glycosylation reactions were repeated twice. (a) Acidic wash and glycosylation 1; (b) acidic wash and glycosylation 2; (c) acidic wash and glycosylation 3; (d) Fmoc deprotection; (e) Nap deprotection; (f) UV cleavage (305 nm); (g) NaOMe, MeOH, 40°C; and (h) Pd/C, H2, MeOH/ethyl acetate/AcOH (30:4:1). Chem 2017 2, 114-124DOI: (10.1016/j.chempr.2016.12.004) Copyright © 2017 Elsevier Inc. Terms and Conditions

Scheme 2 AGA of Sulfated LacNAc and KS Oligosaccharides Reagents and conditions: each glycosylation reaction was executed twice. (a) Acidic wash and glycosylation 1; (b) acidic wash and glycosylation 2; (c) Fmoc deprotection; (d) capping; (e) Nap deprotection; (f) Lev deprotection; (g) sulfation; (h) UV cleavage (305 nm); (i) NaOMe, MeOH, 40°C; and (j) Pd/C, H2, MeOH/triple-distilled water/AcOH (15:15:1). Chem 2017 2, 114-124DOI: (10.1016/j.chempr.2016.12.004) Copyright © 2017 Elsevier Inc. Terms and Conditions