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: 8168929
• 负责人: JAMES H. PRESTEGARD
• 金额: $ 0.17万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-10 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168929
• 关键词: Address; Adopted; Affinity; Affinity Chromatography; Antibodies; Binding; Chemical Structure; Chemicals; Computer Retrieval of Information on Scientific Projects Database; Epitopes; Funding; Grant; Heparitin Sulfate; Immunoglobulin Fragments; Institution; Knowledge; Laboratories; Ligands; Molecular Sieve Chromatography; Oligosaccharides; Pattern; Phage Display; Research; Research Personnel; Resources; Source; Staining method; Stains; Structure-Activity Relationship; Technology; Tissues; United States National Institutes of Health; analog; chemical synthesis; sulfation

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. 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.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 Van Kuppevelt实验室采用噬菌体展示技术生成抗HS单链可变片段抗体，该抗体选择性识别HS寡糖基序。 重要的是，许多抗体显示出各种组织切片的独特染色模式，并且能够区分健康组织和患病组织，后者包括肿瘤组织和肾病理组织。 然而，抗体技术的一个主要限制是缺乏对抗体识别的 HS 表位硫酸化模式的了解。预计 BTBR 技术可以解决这一重要缺陷。该资源将开发的综合方法将用于确定噬菌体展示衍生的单链抗体的配体需求。 在这种方法中，HS 将通过化学和/或酶促方法部分片段化，并通过尺寸排阻色谱和 SAX 分级所得寡糖混合物。 通过亲和纯化分离与抗体高亲和力结合的片段，并通过质谱方法确定化合物的化学结构。 假定的配体和结构类似物将通过模块化方法制备，所得化合物将用于建立结构活性关系（SAR）。 此外，推定的寡糖配体将通过计算方法建立，所得化合物也将进行化学合成。