Translating antibody-binding peptides into peptoid ligands with improved affinity and stability.
Translating antibody-binding peptides into peptoid ligands with improved affinity and stability.
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将抗体结合肽转化为具有改善的亲和力和稳定性的类肽配体。
DOI：
10.1016/j.chroma.2019.05.047
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发表时间：
2019
期刊：
影响因子：
--
通讯作者：
S. Menegatti
中科院分区：
文献类型：
--
作者：
Tee Bordelon;B. Bobay;Andrew J. Murphy;H. Reese;C. Shanahan;Fuad Odeh;A. Broussard;C. Kormos;S. Menegatti
大量的蛋白质结合肽被发现并被用作药物、诊断试剂和亲和配体。然而，最近，肽模拟物由于其优异的生物识别活性和更高的生化稳定性而被提出作为肽的有价值的替代品。这就需要开发一种策略，将已知的蛋白质结合肽翻译成具有相当或更好亲和力的类肽类似物。本研究提出了一种利用igg结合肽HWRGWV作为参考序列的翻译途径。通过调整含有类似天然和非天然氨基酸的官能团的残基的数量和序列排列，合成了HWRGWV的肽类类似物。这些变异最初通过非竞争模式下的igg结合试验筛选，以选择候选配体。一组选择的类肽通过硅对接到人类IgG1、IgG2、IgG3和igg4亚类晶体结构上的推定结合位点上进行了研究，返回的预测结合能值与结合数据非常吻合。选择的肽类PL-16和PL-22进一步通过结合等温线分析进行表征，以确定最大容量（Qmax≈48-57 mg IgG / mL吸附剂）和固相结合强度（KD≈5.4-7.8 10−7M）。吸附剂PL-16-Workbeads和PL-22-Workbeads用于从添加牛血清的细胞培养上清中纯化人IgG，在优化的结合和洗脱条件下，IgG的回收率高达85%，纯度高达98%。这两种肽类配体也被证明对蛋白水解酶和强碱性试剂是稳定的。总的来说，这些研究形成了一种指导同源肽配体的肽变体设计的方法，并有助于解决尽管结构相似，但肽到肽的翻译所面临的挑战。
A great number of protein-binding peptides are known and utilized as drugs, diagnostic reagents, and affinity ligands. Recently, however, peptide mimetics have been proposed as valuable alternative to peptides by virtue of their excellent biorecognition activity and higher biochemical stability. This poses the need to develop a strategy for translating known protein-binding peptides into peptoid analogues with comparable or better affinity. This work proposes a route for translation utilizing the IgG-binding peptide HWRGWV as reference sequence. An ensemble of peptoid analogues of HWRGWV were produced by adjusting the number and sequence arrangement of residues containing functional groups that resemble both natural and non-natural amino acids. The variants were initially screenedviaIgG binding tests in non-competitive mode to select candidate ligands. A set of selected peptoids were studiedin silicoby docking onto putative binding sites identified on the crystal structures of human IgG1, IgG2, IgG3, and IgG4subclasses, returning values of predicted binding energy that aligned well with the binding data. Selected peptoids PL-16 and PL-22 were further characterized by binding isotherm analysis to determine maximum capacity (Qmax˜ 48–57 mg of IgG per mL of adsorbent) and binding strength on solid phase (KD˜ 5.4–7.8 10−7M). Adsorbents PL-16-Workbeads and PL-22-Workbeads were used for purifying human IgG from a cell culture supernatant added with bovine serum, affording high values of IgG recovery (up to 85%) and purity (up to 98%) under optimized binding and elution conditions. Both peptoid ligands also proved to be stable against proteolytic enzymes and strong alkaline agents. Collectively, these studies form a method guiding the design of peptoid variants of cognate peptide ligands, and help addressing the challenges that, despite the structural similarity, the peptide-to-peptoid translation presents.
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DOI：
10.1073/pnas.95.8.4303
发表时间：
1998-04-14
影响因子：
11.1
作者：
Kirshenbaum, K;Barron, AE;Zuckermann, RN
通讯作者：
Zuckermann, RN
影响因子：
2.9
作者：
Johnson, Quentin R.;Lindsay, Richard J.;Nellas, Ricky B.;Fernandez, Elias J.;Shen, Tongye
通讯作者：
Shen, Tongye
影响因子：
7.7
作者：
Scott, Richard W.;DeGrado, William F.;Tew, Gregory N.
通讯作者：
Tew, Gregory N.
影响因子：
4.4
作者：
Proc JL;Kuzyk MA;Hardie DB;Yang J;Smith DS;Jackson AM;Parker CE;Borchers CH
通讯作者：
Borchers CH
调节淀粉样蛋白-β 聚集：肽侧链布局和手性的影响。
影响因子：
3.5
作者：
Turner,JPhillip;Chastain,ShelbyE;Park,Dongwon;Moss,MelissaA;Servoss,ShannonL
通讯作者：
Servoss,ShannonL