Modular synthesis of N-glycans and arrays for the hetero-ligand binding analysis of HIV antibodies

Shivatare, Sachin S.; Chang, Shih-Huang; Tsai, Tsung-I; Tseng, Shih–Ya; Shivatare, Vidya S.; Lin, Yih‐Shyan; Cheng, Yang‐Yu; Ren, Chien‐Tai; Lee, Chang-Chun David; Pawar, Sujeet; Tsai, Charng‐Sheng; Shih, Han C.; Zeng, Yi‐Fang; Liang, Chi-Hui; Kwong, Peter D.; Burton, Dennis R.; Wu, Chung‐Yi; Wong, Chi‐Huey

doi:10.1038/nchem.2463

Cited by 98 publications

(65 citation statements)

References 61 publications

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“…Abs were assessed for glycan reactivity with amine functional glycans printed onto NHS-activated glass slides as previously described (Shivatare et al, 2016). Briefly, amine-functionalized glycans were printed in replicates of three onto NHS-activated glass slides at a concentration of 100 mM.…”

Section: Star+methodsmentioning

confidence: 99%

Glycans Function as Anchors for Antibodies and Help Drive HIV Broadly Neutralizing Antibody Development

Andrabi

Liang

et al. 2017

Immunity

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Summary Apex broadly neutralizing HIV antibodies (bnAbs) recognize glycans and protein surface close to the 3-fold axis of the envelope (Env) trimer and are among the most potent and broad Abs described. The evolution of apex bnAbs from one donor (CAP256) has been studied in detail and many Abs at different stages of maturation have been described. Using diverse engineering tools, we investigated the involvement of glycan recognition in the development of the CAP256.VRC26 Ab lineage. We found that sialic acid-bearing glycans were recognized by germline-encoded and somatically mutated residues on the Ab heavy chain. This recognition provided an “anchor” for the Abs as the core protein epitope varies, prevented complete neutralization escape, and eventually led to broadening of the response. These findings illustrate how glycan-specific maturation enables a human Ab to cope with pathogen escape mechanisms and will aid in optimization of immunization strategies to induce V2 apex bnAb responses.

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Section: Star+methodsmentioning

confidence: 99%

Glycans Function as Anchors for Antibodies and Help Drive HIV Broadly Neutralizing Antibody Development

Andrabi

Liang

et al. 2017

Immunity

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“…If the LacNAc was modified with α1–3Fuc to form Le x , no α2–3-sialylation was detected, consistent with the previous reports. 33,48 When di-LacNAc was in the acceptor ( BA-24 ), within 45 min, the percentage conversion of α2–3-sialylation was 96%, much higher compared to single LacNAc in BA-02 . When the inner LacNAc in di-LacNAc was converted to Le x , the α2–3-sialylation efficiency of the outer LacNAc was even higher than di-LacNAc acceptor, reaching 99%.…”

Section: Resultsmentioning

confidence: 99%

Identification of the binding roles of terminal and internal glycan epitopes using enzymatically synthesized N-glycans containing tandem epitopes

Liu

et al. 2016

Org. Biomol. Chem.

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Glycans play diverse roles in a wide range of biological processes. Research on glycan-binding events is essential for learning their biological and pathological functions. However, the functions of terminal and internal glycan epitopes exhibited during binding with glycan-binding proteins (GBPs) and/or viruses need to be further identified. Therefore, a focused library of 36 biantennary asparagine (Asn)-linked glycans with some presenting tandem glycan epitopes was synthesized via a combined Core Isolation/Enzymatic Extension (CIEE) and one-pot multienzyme (OPME) synthetic strategy. These N-glycans include those containing a terminal sialyl N-acetyllactosamine (LacNAc), sialyl Lewis x (sLex) and Siaα2–8-Siaα2–3/6-R structures with N-acetylneuraminic acid (Neu5Ac) or N-glycolylneuraminic acid (Neu5Gc) sialic acid form, LacNAc, Lewis x (Lex), α-Gal, and Galα1–3-Lex; and tandem epitopes including α-Gal, Lex, Galα1–3-Lex, LacNAc, and sialyl LacNAc, presented with an internal sialyl LacNAc or 1–2 repeats of an internal LacNAc or Lex component. They were synthesized in milligram-scale, purified to over 98% purity, and used to prepare a glycan microarray. Binding studies using selected plant lectins, antibodies, and viruses demonstrated, for the first time, that when interpreting the binding between glycans and GBPs/viruses, not only the structure of the terminal glycan epitopes, but also the internal epitopes and/or modifications of terminal epitopes needs to be taken into account.

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“…A mucin 1 (MUC1) Oglycopeptide array could identify glycopeptide epitopes of cancer-specific antibodies [121], and an Nglycan-derived array successfully determined the glycan-binding specificity of anti-HIV antibodies. In the latter case, enzymatically produced N-glycans including high-mannose, hybrid, and other complextype N-linked oligosaccharides were displayed on an aluminiumoxide-coated glass slide [122]. Due to their complex and heterogeneous nature, the development of GAG-based arrays has been hampered.…”

Section: Large-scale Approaches To Study Early Virus-host Cell Interamentioning

confidence: 99%

Glycomics and Proteomics Approaches to Investigate Early Adenovirus–Host Cell Interactions

Laßwitz

Chandra

Arnberg

2018

Journal of Molecular Biology

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Adenoviruses as most viruses rely on glycan and protein interactions to attach to and enter susceptible host cells. The Adenoviridae family comprises more than 80 human types and they differ in their attachment factor and receptor usage, which likely contributes to the diverse tropism of the different types. In the past years, methods to systematically identify glycan and protein interactions have advanced. In particular sensitivity, speed and coverage of mass spectrometric analyses allow for high-throughput identification of glycans and peptides separated by liquid chromatography. Also, developments in glycan microarray technologies have led to targeted, high-throughput screening and identification of glycan-based receptors. The mapping of cell surface interactions of the diverse adenovirus types has implications for cell, tissue, and species tropism as well as drug development. Here we review known adenovirus interactions with glycan- and protein-based receptors, as well as glycomics and proteomics strategies to identify yet elusive virus receptors and attachment factors. We finally discuss challenges, bottlenecks, and future research directions in the field of non-enveloped virus entry into host cells.

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Modular synthesis of N-glycans and arrays for the hetero-ligand binding analysis of HIV antibodies

Cited by 98 publications

References 61 publications

Glycans Function as Anchors for Antibodies and Help Drive HIV Broadly Neutralizing Antibody Development

Glycans Function as Anchors for Antibodies and Help Drive HIV Broadly Neutralizing Antibody Development

Identification of the binding roles of terminal and internal glycan epitopes using enzymatically synthesized N-glycans containing tandem epitopes

Glycomics and Proteomics Approaches to Investigate Early Adenovirus–Host Cell Interactions

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