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Large scale generation of anti-zebrafish monoclonal antibodies

大规模产生抗斑马鱼单克隆抗体

基本信息

批准号：
8304115
负责人：
Gavin James Wright
金额：
$ 18.4万
依托单位：
SANGER INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-07-01 至 2013-12-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8304115
关键词：
Alkaline Phosphatase Animal Model Antibodies Antibody Affinity Axon Biological Process Biotin Categories Cell Polarity Cell Surface Proteins Cell surface Cells Clonality Cloning Communities Cryoultramicrotomy Data Development Disease Embryo Ensure Ephrins Epitopes Family Formalin Gene Targeting Generations Goals Hybridomas Hybrids Immunization In Situ Hybridization Label Laboratories Laboratory Organism Lead Libraries Location Methods Monoclonal Antibodies Names Neurons Pattern Plasmids Process Protein Microchips Proteins Protocols documentation Reagent Recombinants Research Research Personnel Research Proposals Resistance Resources Site Staging Surface Technology Testing Therapeutic Intervention Tissues Transgenic Animals Zebrafish Zebrafish Proteins base cell type design gene function genetic resource in vivo insight intracellular protein transport junctional adhesion molecule morphogens mutant neuronal cell body novel novel strategies protein expression protein function public health relevance symposium tool

项目摘要

DESCRIPTION (provided by applicant): This proposal outlines how a unique resource of zebrafish cell surface and secreted proteins will be used in conjunction with a novel method of antibody selection to create a resource of 200 new zebrafish monoclonal antibody reagents. The antibody targets have been carefully selected primarily from their known in situ hybridization patterns so as to be of maximum benefit to the zebrafish community. Targets include: new cellular markers and proteins whose subcellular localization is important for function such as morphogens, polarity proteins and neural cell surface proteins. The throughput required to select this large number of antibodies will be achieved by using a novel pooling immunization strategy, antibody selection using protein microarrays and the recombinant cloning of antibody V-regions. The antibody selection process will identify antibodies that are non-crossreactive with proteins from the same family and recognize formalin-resistant epitopes. Finally, each antibody will be tested on whole mount embryos and cryosections and will be freely and widely distributed. These antibodies, when used in conjunction with the large genetic resources available to zebrafish researchers, will allow new mechanistic insights into protein function which may eventually be used to design novel strategies for therapeutic intervention. PUBLIC HEALTH RELEVANCE: The zebrafish is an increasingly popular laboratory organism with many experimental advantages that biologists exploit to discover the detailed processes that underlie vertebrate development and disease. Zebrafish research, however, is limited by a paucity of good antibody reagents that are essential tools for determining the precise details of biological processes. This research proposal is aimed at providing the zebrafish community with a major resource of new antibody reagents.

描述（由申请人提供）：本提案概述了如何将斑马鱼细胞表面和分泌蛋白的独特资源与抗体选择的新方法结合使用，以创建200种新斑马鱼单克隆抗体试剂的资源。主要从已知的原位杂交模式中仔细选择抗体靶标，以便对斑马鱼群落具有最大益处。具体目标包括：新的细胞标记物和蛋白质，其亚细胞定位对于功能是重要的，例如形态发生蛋白、极性蛋白和神经细胞表面蛋白。选择大量抗体所需的通量将通过使用新的合并免疫策略、使用蛋白质微阵列的抗体选择和抗体V区的重组克隆来实现。抗体选择过程将鉴定与来自相同家族的蛋白质无交叉反应性的抗体，并识别福尔马林抗性表位。最后，每种抗体将在整个胚胎和冷冻切片上进行测试，并将自由和广泛分布。当这些抗体与斑马鱼研究人员可用的大量遗传资源结合使用时，将允许对蛋白质功能进行新的机制见解，最终可能用于设计新的治疗干预策略。公共卫生关系：斑马鱼是一种越来越受欢迎的实验室生物，具有许多实验优势，生物学家可以利用这些优势来发现脊椎动物发育和疾病的详细过程。然而，斑马鱼的研究受到缺乏良好抗体试剂的限制，这些抗体试剂是确定生物过程精确细节的必要工具。这项研究计划旨在为斑马鱼社区提供新的抗体试剂的主要资源。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A panel of recombinant monoclonal antibodies against zebrafish neural receptors and secreted proteins suitable for wholemount immunostaining.

一组针对斑马鱼神经受体和分泌蛋白的重组单克隆抗体，适合整体免疫染色。

DOI：
10.1016/j.bbrc.2014.11.123
发表时间：
2015
期刊：
Biochemical and biophysical research communications
影响因子：
3.1
作者：
Staudt,Nicole;Müller-Sienerth,Nicole;Fane-Dremucheva,Alla;Yusaf,ShahnazP;Millrine,David;Wright,GavinJ
通讯作者：
Wright,GavinJ

A rapid and scalable method for selecting recombinant mouse monoclonal antibodies.