HUMANIZED ANTITUMOR BINDING SITES--FRAMEWORK ENGINEERING

人源化抗肿瘤结合位点--框架工程

• 批准号: 3492525
• 负责人: JAMES S. HUSTON
• 金额: $ 5万
• 依托单位: CREATIVE BIOMOLECULES, INC.
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-06-01 至 1992-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3492525
• 关键词: antibody; antigens; bacteria; chemical binding; chemical models; circular dichroism; digoxin; fusion gene; gene expression; genetic manipulation; immunoglobulin genes; immunoglobulin structure; lysozyme; molecular cloning; monoclonal antibody; protein engineering; protein folding; protein reconstitution; protein structure function; secretion

DESCRIPTION: (Adapted from the Applicant's Abstract) Antibody variable (v) regions each comprise 3 complementary determining regions (CDRs) that form the antigen binding site and 4 framework regions (FRs) that form the scaffold supporting the binding site. These segments are interspersed sequentially in each V region to form FR1-CDR-1 FR2-CDR-2-FR3-CDR3-FR4, with the whole binding site requiring VH and VL. This proposal explores the modification of the FR structure for the improvement of immunochemical and solution properties of minimal antibody binding sites, consisting of single-chain Fv (sFv, comprising VH-linker-VL) or single domain V regions. In the folded conformation of each V domain, FRs form a barrel of beta pleated sheet and the CDRs form loops that connect the inner FRs to form the antigen binding site. Fr sequences from human antibodies can be fused to CDRs (a process termed humanization (which eliminates much of the antigenicity contributed by murine antigenic sites. This proposal seeks to develop advanced FR designs to be used in making hybrid sFv or variable domains, having improved stability/solubility and reduced sFv immunogenicity. The investigators will produce a humanized 26-10 antidigoxin sFv and a murine-murine hybrid sFv with an antilysozyme binding site grafted onto the 26-10 framework. In addition, the investigators will explore minimal binding sites comprising only the 26-10 VH domain, as a monomer or single-chain homodimer. Finally, they will analyze an engineered human framework which retains only simple -gly-gly-ser-gly-gly-loops to connect FRs. This nonfunctional "framework only" molecule could be a cornerstone for rapid development of very well behaved biosynthetic antibody binding sites. It would allow any FR superstructure to be directly analyzed and optimized without the perturbation of a particular binding site; specific CDRs would be reinserted for applications to tumor imaging, therapy, etc.

描述：（改编自申请人摘要）抗体变量(v)

项目成果

Symmetry of Fv architecture is conducive to grafting a second antibody binding site in the Fv region.

Fv结构的对称性有利于在Fv区移植第二抗体结合位点。

• DOI: 10.1016/s0006-3495(96)79398-0
• 发表时间: 1996
• 期刊: Biophysical journal.
• 作者: Keck,PC;Huston,JS
• 通讯作者: Huston,JS