EPITOPE SCANNING USING PROTEIN SEMISYNTHESIS

使用蛋白质半合成进行表位扫描

• 批准号: 2023427
• 负责人: Ashley J Birkett
• 金额: $ 10万
• 依托单位: IMMUNE COMPLEX CORPORATION
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-02-01 至 1998-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2023427
• 关键词: Escherichia coli; antigen presentation; bacterial proteins; biotechnology; hepatitis B virus group; nucleocapsid; peptide chemical synthesis; peptide structure; protein sequence; synthetic peptide; transfection /expression vector; transposon /insertion element

DESCRIPTION: (Adapted from applicant's abstract) Recent advances in the chemoselective ligation of peptides has greatly increased the range of useful proteins that can be produced in this manner. The adaptation of this procedure to facilitate the internal insertion of designer synthetic peptides between recombinantly derived protein fragments will be studied using a particular carrier protein. This protein spontaneously assembles into ordered, soluble, 27 nm particles containing 180 subunits. This structure has been demonstrated to be a superior carrier for exogenous, pathogen derived epitopes. The chemoselective insertion of peptide cassettes between recombinant N- and C-terminal fragments of the particulate carrier will be investigated to determine the feasibility of this directional, three-piece ligation procedure for generating novel fusion proteins harboring synthetic sequences. The physical, chemical and immunological integrity of the semisynthetic particles will be examined. A critical comparison of these data with those acquired for the entirely recombinant equivalents will enable an assessment of the utility of this approach. This work is expected to yield a versatile method for the introduction of B- and T-cell sites as well as novel amino acids or other chemical entities into a proven carrier protein that dramatically enhances immune responses to inserted sequences and conjugated haptens. Rapid production of analytical quantities of hybrid proteins will enable epitope scanning to identify useful antibody reactivities.

描述：(改编自申请人的摘要)最近在 多肽的化学选择性连接大大增加了其作用范围。 可以用这种方式产生的有用的蛋白质。改编自《纽约时报》 本程序便于设计者在内部插入合成 重组衍生的蛋白质片段之间的多肽将被研究 使用一种特殊的载体蛋白。这种蛋白质会自发地组装 形成有序的、可溶的、包含180个亚基的27 nm颗粒。这 结构已被证明是外源， 病原体衍生的表位。多肽的化学选择性插入 重组的N-末端和C-末端片段之间的盒带 将对颗粒载体进行调查，以确定 这种定向的、三件式的结扎程序产生了新的 含有合成序列的融合蛋白。物理的、化学的 半合成粒子的免疫学完整性将是 检查过了。将这些数据与为获得的数据进行关键比较 完全重组的等价物将使评估 这种方法的实用性。预计这项工作将产生一种通用的 引入B细胞和T细胞位点的方法以及新的氨基 酸或其他化学物质转化为已证实的载体蛋白 显著增强对插入序列的免疫反应，并 共轭半抗原。杂交种分析量的快速生产 蛋白质将使表位扫描能够识别有用的抗体 反应性。