Scholarlink

<p>The study of natural glycans is fundamental to modern glycomics; however, the functional analysis of these glycans is limited by their low natural abundance, structural heterogeneity, and the absence of efficient preparative-scale isolation methods. Here, we present a streamlined approach for the gram-scale production of complex natural N-glycans in their reducing form by combining oxidative release of natural glycans (ORNG) using household bleach, an innovative cleavable tag chemistry, and optimized multi-dimensional chromatography. Our ORNG process releases N-glycans from kilogram-scale of natural sources containing various glycoproteins, which can be efficiently attached with a 4-aminobenzoate tag, facilitating the selective separation of these glycans from other biomolecules. The tagged glycans can be purified via dry-load HILIC flash chromatography at gram scale followed by reverse-phase HPLC. Treatment with Oxone quantitatively cleaves the tag, regenerating the free reducing N-glycans in high yield. This method provides an accessible, gram-scale source of complex N-glycans, including complex asymmetric structures that are challenging to synthesize through conventional chemoenzymatic approaches. We believe this approach represents a versatile strategy for acquiring complex natural glycans, opening new avenues for the functional exploration of N-glycans in glycoscience.</p><p></p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Streamlined gram-scale natural N-glycan production using reversible tagging after oxidative release of natural glycans

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2025年4月4日

查看全文
  • Qing Zhang,
  • Yi Lasanajak,
  • Menxin Yan,
  • Golden Chen,
  • Xuezheng Song

摘要

The study of natural glycans is fundamental to modern glycomics; however, the functional analysis of these glycans is limited by their low natural abundance, structural heterogeneity, and the absence of efficient preparative-scale isolation methods. Here, we present a streamlined approach for the gram-scale production of complex natural N-glycans in their reducing form by combining oxidative release of natural glycans (ORNG) using household bleach, an innovative cleavable tag chemistry, and optimized multi-dimensional chromatography. Our ORNG process releases N-glycans from kilogram-scale of natural sources containing various glycoproteins, which can be efficiently attached with a 4-aminobenzoate tag, facilitating the selective separation of these glycans from other biomolecules. The tagged glycans can be purified via dry-load HILIC flash chromatography at gram scale followed by reverse-phase HPLC. Treatment with Oxone quantitatively cleaves the tag, regenerating the free reducing N-glycans in high yield. This method provides an accessible, gram-scale source of complex N-glycans, including complex asymmetric structures that are challenging to synthesize through conventional chemoenzymatic approaches. We believe this approach represents a versatile strategy for acquiring complex natural glycans, opening new avenues for the functional exploration of N-glycans in glycoscience.