A rationally designed agonist antibody fragment that functionally mimics thrombopoietin
A rationally designed agonist antibody fragment that functionally mimics thrombopoietin
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DOI：
10.1073/pnas.0602658103
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发表时间：
2006-09-26
影响因子：
11.1
通讯作者：
Bowdish, Katherine S.
中科院分区：
文献类型：
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作者：
Frederickson, Shana;Renshaw, Mark W.;Bowdish, Katherine S.
通过合理设计，产生模拟血小板生成素（TPO）的抗体片段（Fab）。将具有cMpl受体结合能力的肽移植到全人Fab支架的不同互补决定区中。通过噬菌体展示和基于细胞的淘选优化肽的功能呈递。测定含有两种移植肽的选择抗体和片段在体外刺激cMpl受体的能力。几种候选物在体外cMpl受体信号传导报告基因测定中表现出激动剂活性，包括Fab59，估计其与TPO等效。Fab59还能够有效地刺激正常小鼠的血小板生成。这些合理设计的模拟Fab可以提供血小板减少症的治疗干预，同时避免内源性TPO的中和抗体的潜在产生。此外，本研究证明了一种方法，通过该方法可以将具有结合活性的短寿命线性肽转化为能够激活细胞表面受体的更稳定和有效的激动剂。
By using rational design, antibody fragments (Fabs) that mimic thrombopoietin (TPO) were created. A peptide with cMpl receptor-binding capability was grafted into different complementarity-determining regions of a fully human Fab scaffold. Functional presentation of the peptide was optimized by using phage display and cell-based panning. Select antibodies and fragments containing two grafted peptides were assayed for their ability to stimulate the cMpl receptor in vitro. Several candidates demonstrated agonist activity in an in vitro cMpl receptor signaling reporter assay, including Fab59, which was estimated to be equipotent to TPO. Fab59 additionally was able to effectively stimulate platelet production in normal mice. These rationally designed mimetic Fabs may provide a therapeutic intervention for thrombocytopenia while avoiding the potential generation of neutralizing antibodies to endogenous TPO. Furthermore, this study demonstrates a method by which short-lived linear peptides with binding activity may be converted to more stable and potent agonists capable of activating cell surface receptors.