HEPARAN SULFATE LIGAND REQUIREMENTS OF PHAGE DISPLAY ANTIBODIES

噬菌体展示抗体的硫酸乙酰肝素配体要求

• 批准号: 8361820
• 负责人: JAMES H. PRESTEGARD
• 金额: $ 0.18万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-01 至 2012-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361820
• 关键词: Address; Adopted; Affinity; Affinity Chromatography; Antibodies; Binding; Chemical Structure; Chemicals; Epitopes; Funding; Grant; Heparitin Sulfate; Immunoglobulin Fragments; Knowledge; Laboratories; Ligands; Molecular Sieve Chromatography; National Center for Research Resources; Oligosaccharides; Pattern; Phage Display; Principal Investigator; Research; Research Infrastructure; Resources; Source; Staining method; Stains; Structure-Activity Relationship; Technology; Tissues; United States National Institutes of Health; analog; chemical synthesis; cost; sulfation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The Van Kuppevelt laboratory has adopted phage display technology to generate anti-HS single chain variable fragment antibodies, which selectively recognize HS oligosaccharide motifs. Importantly, many of the antibodies show unique staining patterns of various tissue sections and have an ability to distinguish between healthy and diseased tissue, the latter including tumorous and nephropathological tissue. However, a major limitation of the antibody technology is a lack of knowledge of sulfation patterns of HS epitopes that are recognized by the antibodies. It is to be expected that the BTBR technology can address this important deficiency. The integrated approach that will be developed by this resource will be employed to identify ligand requirements of the phage display derived single chain antibodies. In this approach, HS will be partially fragmented by chemical and/or enzymatic approaches and the resulting mixtures of oligosaccharides fractionated by size exclusion chromatography and SAX. Fragments that bind with high affinity to an antibody will be isolated by affinity purification and the chemical structures of the compounds determined by mass spectroscopic approaches. Putative ligands and structural analogs will be prepared by a modular approach and the resulting compounds will be employed to establish structure activity relationships (SAR). In addition, putative oligosaccharide ligand will be established by computational approaches and the resulting compounds will also be subjected to chemical synthesis.

该副本是利用资源的众多研究子项目之一 由NIH/NCRR资助的中心赠款提供。对该子弹的主要支持 而且，副投影的主要研究员可能是其他来源提供的 包括其他NIH来源。 列出的总费用可能 代表subproject使用的中心基础架构的估计量， NCRR赠款不直接向子弹或副本人员提供的直接资金。 Van Kuppevelt实验室采用了噬菌体显示技术来生成抗HS单链可变片段抗体，该抗体有选择地识别HS寡糖基序。 重要的是，许多抗体显示了各种组织切片的独特染色模式，并且具有区分健康和患病组织的能力，后者包括肿瘤和肾病组织。 但是，抗体技术的主要局限性是缺乏对抗体识别的HS表位的硫酸化模式的了解。可以预期，BTBR技术可以解决这一重要缺陷。将采用该资源开发的集成方法来确定噬菌体显示派生的单链抗体的配体要求。 在这种方法中，HS将通过化学和/或酶促方法部分碎片，以及由尺寸排除色谱和萨克斯（SAX）分级分离的寡糖的混合物。 与抗体高亲和力结合的片段将通过亲和力纯化和通过质谱方法确定的化合物的化学结构分离。 假定的配体和结构类似物将通过模块化方法制备，并将采用所得化合物来建立结构活动关系（SAR）。 此外，将通过计算方法建立推定的寡糖配体，所得化合物也将经过化学合成。