Engineered synthetic polymer nanoparticles as IgG affinity ligands.
Engineered synthetic polymer nanoparticles as IgG affinity ligands.
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DOI：
10.1021/ja303612d
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发表时间：
2012-09-26
影响因子：
15
通讯作者：
Shea, Kenneth J.
中科院分区：
文献类型：
--
作者：
Lee, Shih-Hui;Hoshino, Yu;Randall, Arlo;Zeng, Zhiyang;Baldi, Piere;Doong, Ruey-an;Shea, Kenneth J.
描述了一种制备非生物蛋白亲和配体的方法。亲和配体是一种合成聚合物水凝胶纳米颗粒（NP），其官能团与目标蛋白的表面呈现相辅相成。迭代过程通过优化功能单体的组成和比例来提高亲和性。由于聚合物NPs是由动力学驱动的过程形成的，因此聚合物链中功能单体的顺序不受控制；饲料中单体的化学计量只能调整平均组成。为了弥补这一点，水凝胶NP轻度交联，从而在亚毫秒的时间尺度上产生链的灵活性，从而使聚合物能够“映射”到具有互补功能的蛋白质表面。在这项研究中，我们报道了一种轻交联（2%）n -异丙基丙烯酰胺（NIPAm）合成聚合物NP (50~65 nm)，包含疏水性和羧酸基团，与IgG的Fc片段具有高亲和力。NP与IgG结合的亲和力和数量与pH值有关。水凝胶NP在pH为5.5时抑制蛋白A与Fc结构域的结合，而在pH为7.3时则没有。计算分析用于确定潜在的np -蛋白相互作用位点。候选者包括一个NP结合域，在pH为5.5时与蛋白a - fc结合域重叠。计算分析支持抑制实验结果，并归因于组氨酸残基带电状态的差异。在pH 5.5下，NP （3.5~8.5 nM）对Fc结构域的亲和力与蛋白A在pH 7下的亲和力相当。这些结果表明，工程合成的聚合物NPs可以与大型生物大分子的特定区域具有内在亲和力。
A process for the preparation of an abiotic protein affinity ligand is described. The affinity ligand, a synthetic polymer hydrogel nanoparticle (NP), is formulated with functional groups complementary to the surface presentation of the target protein. An iterative process is used to improve affinity by optimizing the composition and proportion of functional monomers. Since the polymer NPs are formed by a kinetically driven process, the sequence of functional monomers in the polymer chain is not controlled; only the average composition can be adjusted by the stoichiometry of the monomers in the feed. To compensate for this the hydrogel NP is lightly crosslinked resulting in chain flexibility that takes place on a sub millisecond time scale allowing the polymer to “map” onto a protein surface with complementary functionality. In this study, we report a lightly crosslinked (2%) N-isopropyl acrylamide (NIPAm) synthetic polymer NP (50~65 nm) incorporating hydrophobic and carboxylate groups, binds with high affinity to the Fc fragment of IgG. The affinity and amount of NP bound to IgG is pH dependent. The hydrogel NP inhibits protein A binding to the Fc domain at pH 5.5, but not at pH 7.3. A computational analysis was used to identify potential NP-protein interaction sites. Candidates include a NP binding domain that overlaps with the protein A-Fc binding domain at pH 5.5. The computational analysis supports the inhibition experimental results and is attributed to the difference in the charged state of histidine residues. Affinity of the NP (3.5~8.5 nM) to the Fc domain at pH 5.5 is comparable to protein A at pH 7. These results establish that engineered synthetic polymer NPs can be formulated with an intrinsic affinity to a specific domain of a large biomacromolecule.
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影响因子：
16.6
作者：
Phillips, Ronnie L.;Miranda, Oscar R.;Bunz, Uwe H. F.
通讯作者：
Bunz, Uwe H. F.
影响因子：
4.1
作者：
Lin, FY;Chen, CS;Yamamoto, S
通讯作者：
Yamamoto, S
影响因子：
7.4
作者：
Ozeki, Tomomitsu;Morita, Mizuki;Okahata, Yoshio
通讯作者：
Okahata, Yoshio
影响因子：
10.8
作者：
Messerschmidt, Sylvia K. E.;Musyanovych, Anna;Kontermann, Roland E.
通讯作者：
Kontermann, Roland E.
影响因子：
2.9
作者：
RAGHAVAN, M;BONAGURA, VR;BJORKMAN, PJ
通讯作者：
BJORKMAN, PJ