A rapid and scalable method for selecting recombinant mouse monoclonal antibodies.
A rapid and scalable method for selecting recombinant mouse monoclonal antibodies.
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DOI：
10.1186/1741-7007-8-76
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发表时间：
2010-06-04
期刊：
影响因子：
5.4
通讯作者：
Wright GJ
中科院分区：
文献类型：
--
作者：
Crosnier C;Staudt N;Wright GJ
对于细胞和发育生物学家来说，具有高亲和力和选择性的单克隆抗体对整个固定组织起作用，是有价值的试剂，但分离它们仍然是一个漫长且不可预测的过程。在这里，我们报告了一种快速和可扩展的方法来选择和表达重组小鼠单克隆抗体，这些抗体基本上相当于由亲本IgG同种型杂交瘤分泌的抗体。通过用抗原库免疫小鼠并从少量杂交瘤细胞克隆功能重排的轻链和重链到单个表达质粒中来实现增加的通量。通过免疫与哺乳动物细胞中表达的斑马鱼细胞表面受体蛋白的胞外结构域和福尔马林抗性表位的筛选，我们选择的抗体，给出了预期的染色模式的wholemount固定的斑马鱼胚胎。该方法可用于从一只免疫小鼠中快速筛选出几种高质量的单克隆抗体，并便于它们使用质粒进行分布。
Monoclonal antibodies with high affinity and selectivity that work on wholemount fixed tissues are valuable reagents to the cell and developmental biologist, and yet isolating them remains a long and unpredictable process. Here we report a rapid and scalable method to select and express recombinant mouse monoclonal antibodies that are essentially equivalent to those secreted by parental IgG-isotype hybridomas. Increased throughput was achieved by immunizing mice with pools of antigens and cloning - from small numbers of hybridoma cells - the functionally rearranged light and heavy chains into a single expression plasmid. By immunizing with the ectodomains of zebrafish cell surface receptor proteins expressed in mammalian cells and screening for formalin-resistant epitopes, we selected antibodies that gave expected staining patterns on wholemount fixed zebrafish embryos. This method can be used to quickly select several high quality monoclonal antibodies from a single immunized mouse and facilitates their distribution using plasmids.
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影响因子：
7
作者：
Bushell, K. Mark;Sollner, Christian;Wright, Gavin J.
通讯作者：
Wright, Gavin J.
影响因子：
2.5
作者：
KIMMEL, CB;BALLARD, WW;SCHILLING, TF
通讯作者：
SCHILLING, TF
影响因子：
82.9
作者：
Jin, Aishun;Ozawa, Tatsuhiko;Muraguchi, Atsushi
通讯作者：
Muraguchi, Atsushi
影响因子：
48
作者：
Taussig, Michael J.;Stoevesandt, Oda;Uhlen, Mathias
通讯作者：
Uhlen, Mathias
影响因子：
3.4
作者：
De Masi, F;Chiarella, P;Sawyer, A
通讯作者：
Sawyer, A