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.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 Van Kuppevelt实验室采用噬菌体展示技术制备抗HS单链可变区抗体，选择性识别HS寡糖基序。重要的是，许多抗体在不同的组织切片上显示出独特的染色模式，并具有区分健康组织和疾病组织的能力，后者包括肿瘤组织和肾脏病理组织。然而，抗体技术的一个主要限制是缺乏对抗体识别的HS表位的硫酸盐化模式的了解。可以预期，BTBR技术可以解决这一重要缺陷。这一资源将开发的综合方法将用于鉴定噬菌体展示单链抗体的配基需求。在这种方法中，HS将通过化学和/或酶方法进行部分裂解，得到的低聚糖混合物将通过尺寸排除层析和SAX进行分级。与抗体具有高亲和力的片段将通过亲和纯化和通过质谱学方法确定化合物的化学结构来分离。假定的配体和结构类似物将通过模块化方法制备，所得到的化合物将用于建立结构活性关系(SAR)。此外，假定的寡糖配体将通过计算方法建立，得到的化合物也将经过化学合成。