Design and synthesis of a photocleavable biotin-linker for the photoisolation of ligand-receptor complexes based on the photolysis of 8-quinolinyl sulfonates in aqueous solution
Design and synthesis of a photocleavable biotin-linker for the photoisolation of ligand-receptor complexes based on the photolysis of 8-quinolinyl sulfonates in aqueous solution
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DOI：
10.1016/j.bmc.2009.03.031
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发表时间：
2009-05-01
影响因子：
3.5
通讯作者：
Kageyama, Yoshiyuki
中科院分区：
文献类型：
--
作者：
Aoki, Shin;Matsuo, Nanako;Kageyama, Yoshiyuki
亲和素（Avn）与生物素（Btn）形成强络合物的能力在生化、分析和医学研究中经常被用于生物分子的检测和分离。然而，这种结合几乎是不可逆的，这在分离和纯化完整的生物分子方面构成了一个缺点。我们最近发现芳香或脂肪族磺酸的8-喹啉基酯在中性ph的水溶液中照射300-330 nm时发生光解。在这项工作中，设计并合成了一种生物素-多巴胺（BD）偶联物，该偶联物含有可光裂解的8-喹啉基苯磺酸（QB）连接剂BDQB，用于从Avn-Btn系统中有效回收多巴胺-蛋白（例如抗体）复合物。在avn包被板上，BDQB与一抗多巴胺抗体（小鼠抗多巴胺IgG(1)）通过酶联免疫吸附试验（ELISA）证实，二抗（抗IgG(1)抗体）与辣根过氧化物酶（HRP）偶联。BDQB-IgG(1)复合物经313 nm光照射后，ELISA证实其释放多巴胺- igg(1)复合物。通过SDS-PAGE和Western blot对所获得的光释放多巴胺抗多巴胺IgG(1)复合物进行表征。2009爱思唯尔有限公司版权所有。
The ability of avidin (Avn) to form strong complex with biotin (Btn) is frequently used in the detection and isolation of biomolecules in biochemical, analytical, and medicinal research. The fact that the binding is nealy irreversible, however, constitutes a drawback in term of the isolation and purification of intact biomolecules. We recently found that 8-quinolinyl esters of aromatic or aliphatic sulfonic acids undergo photolysis when irradiated at 300-330 nm in aqueous solution at neutral pH. In this work, a biotin-dopamine (BD) conjugate containing a photocleavable 8-quinolinyl benzenesulfonate (QB) linker, BDQB, was designed and synthesized for use in the efficient recovery of dopamine-protein (e.g., antibody) complexes from an Avn-Btn system. The complexation of BDQB with a primary anti-dopamine antibody (anti-dopamine IgG(1) from mouse) on an Avn-coated plate was confirmed by an enzyme-linked immunosorbent assay ( ELISA) utilizing a secondary antibody (anti-IgG(1) antibody) conjugated with horseradish peroxidase (HRP). Upon the photoirradiation (at 313 nm) of the BDQB-IgG(1) complex, the release of dopamine-IgG(1) complex was confirmed by ELISA. Characterization of the resulting photoreleased dopamine anti-dopamine IgG(1) complex was performed by SDS-PAGE and Western blot. (C) 2009 Elsevier Ltd. All rights reserved.