Facile Conjugation of Biomolecules onto Surfaces via Mussel Adhesive Protein Inspired Coatings.
Facile Conjugation of Biomolecules onto Surfaces via Mussel Adhesive Protein Inspired Coatings.
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DOI：
10.1002/adma.200801222
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发表时间：
2009-01-26
影响因子：
29.4
通讯作者：
Messersmith, Phillip B.
中科院分区：
文献类型：
--
作者：
Lee, Hoeshin;Rho, Junsung;Messersmith, Phillip B.
生物分子在表面上的固定在许多生物和物理科学中都很重要，包括细胞和分子生物学，分析化学，以及应用和跨学科领域，如医学诊断，组织工程和生物过程工程。[1-4]生物分子在表面上的固定策略通常利用非共价或共价反应。非共价方法允许在特定条件下可逆固定生物分子，包括物理吸附和亲和固定。一些广泛适用的例子是（链球菌）亲和素-生物素，硝基乙酸(NTA)-组氨酸和DNA-DNA相互作用。[5-8]相比之下，分子在表面上的共价固定通常依赖于“活性”官能团之间的偶联反应，如n -羟基琥珀酰亚胺（NHS）[9]或马来酰亚胺[10]和伴靶基团，如胺和巯基。对于在水溶液中进行的涉及生物分子的反应，NHS、马来酰亚胺和其他活化基团在储存和反应过程中对水解的敏感性可能导致表面生物偶联效率低。[11,12]在这项研究中，我们报告了一种简单的两步水相方法将生物分子固定在表面上。该方法包括简单地将仿生聚合物薄膜浸涂在衬底上，然后将生物分子偶联到仿生聚合物薄膜上。该方法利用了仿生聚合物薄膜对亲核试剂的潜在反应性，不受水的影响，并允许在pKa的基础上区分亲核试剂。底物首先在碱性多巴胺·HCl溶液（2mg多巴胺·HCl溶解在10 mM Tris缓冲液中，pH为8.5）中浸泡12-18小时，然后涂上一层薄的聚多巴胺薄膜多巴胺是一种含有儿茶酚和胺官能团的生物分子，也高浓度存在于贻贝黏附蛋白中，[14,15]在碱性ph下聚合形成薄的黏附聚多巴胺膜，对胺和巯基表现出潜在的反应性。[13,16]在该方法的第二步中，利用聚多巴胺膜的反应性，通过亲核试剂和聚多巴胺表面之间的反应将生物分子共价固定在表面上（方案1）。
Immobilization of biomolecules onto surfaces is important in many of the biological and physical sciences, including cell and molecular biology, analytical chemistry, and in applied and interdisciplinary fields such as medical diagnostics, tissue engineering, and bioprocess engineering.[1-4] Strategies for biomolecule immobilization onto surfaces generally exploit either noncovalent or covalent reactions. Noncovalent methods allow reversible immobilization of biomolecules under specific conditions, and include physical adsorption and affinity immobilization. Some widely adapted examples are (strep) avidin-biotin, nitriloacetic acid (NTA)-histidine, and DNA-DNA interactions.[5-8] In contrast, covalent immobilization of molecules onto surfaces typically relies on conjugation reactions between ‘active’functional groups, such as N-hydroxysuccinimide (NHS)[9] or maleimide,[10] and companion target moieties, such as amines and sulfhydryls. For reactions involving biomolecules performed in aqueous solvents, susceptibility of NHS, maleimide, and other activating groups to hydrolysis during storage and reaction can lead to low efficiency of surface bioconjugation.[11, 12] In this study, we report a facile two-step aqueous approach to immobilization of biomolecules onto surfaces. The approach involves simple dip-coating of a biomimetic polymer thin film onto a substrate, followed by conjugation of biomolecules to the biomimetic polymer film. The method exploits the latent reactivity of the biomimetic polymer thin film towards nucleophiles, is unaffected by water, and allows for discrimination between nucleophiles on the basis of pKa.Substrates were first coated with a thin adherent polydopamine film by immersion in an alkaline dopamine· HCl solution (2 mg of dopamine· HCl dissolved in 10 mM Tris buffer, pH 8.5) for up to 12-18 h.[13] Dopamine, a biomolecule that contains catechol and amine functional groups, found also in high concentration in mussel adhesive proteins,[14, 15] polymerizes at alkaline pHs to form thin adherent polydopamine films that exhibit latent reactivity toward amine and thiol groups.[13, 16] In the second step of the approach, the reactivity of the polydopamine films is exploited to covalently immobilize biomolecules onto surfaces through a reaction between nucleophiles and the polydopamine surface (Scheme 1).
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DOI：
10.1073/pnas.072685299
发表时间：
2002-04-16
影响因子：
11.1
作者：
Hodneland, CD;Lee, YS;Mrksich, M
通讯作者：
Mrksich, M
影响因子：
4.8
作者：
PAPOV, VV;DIAMOND, TV;WAITE, JH
通讯作者：
WAITE, JH
影响因子：
3.8
作者：
Chen, H;Hsieh, YL
通讯作者：
Hsieh, YL
影响因子：
7.4
作者：
Schmid, EL;Keller, TA;Vogel, H
通讯作者：
Vogel, H
影响因子：
56.9
作者：
Lee, Haeshin;Dellatore, Shara M.;Messersmith, Phillip B.
通讯作者：
Messersmith, Phillip B.