Preparation and Characterization of Molecularly Imprinted Polymeric Nanoparticles for Atrial Natriuretic Peptide (ANP).
Preparation and Characterization of Molecularly Imprinted Polymeric Nanoparticles for Atrial Natriuretic Peptide (ANP).
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DOI：
10.1002/adfm.201100946
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发表时间：
2011-12-06
影响因子：
19
通讯作者：
Mohapatra, Subhra
中科院分区：
文献类型：
--
作者：
Wang, Chunyan;Howell, Mark;Raulji, Payal;Davis, Yvonne;Mohapatra, Subhra
利钠肽受体A （NPRA）是心脏激素心房利钠肽（ANP）的受体，在癌细胞上大量表达，破坏ANP-NPRA信号通路可抑制肿瘤负荷和转移。由于NPRA信号的拮抗剂没有提供可重复的结果，我们推断，一种对ANP具有高选择性和亲和力的合成的ANP中和抗体可以用来调节ANP水平并减弱NPRA信号。本研究以ANP短肽为模板制备了ANP分子印迹聚合物纳米颗粒（MIPNPs），并测定了其结合亲和力和选择性。以ANP短肽NH2-SLRRSS-CONH2为模板，甲基丙烯酸（MAA）和n -异丙基丙烯酰胺（NIPAm）为功能单体，双丙烯酰胺（BIS）为交联剂，采用沉淀聚合法制备了MIPNPs。MIPNPs和非印迹纳米颗粒（NIPNPs）在水中的平均直径分别为215.8±4.6 nm和197.7±3.1 nm。结合等温线分析表明，与NIPNPs相比，MIPNPs对模板肽和ANP具有更高的结合亲和力。经Scatchard分析，其平衡解离常数Kd为7.3 μM，与模板肽的结合容量为106.7 μmol/g； Kd为7.9 μM，与ANP的结合容量为36.0 μmol/g。结合动力学测量表明，MIPNPs在30分钟内达到蛋白质吸附平衡。发现MIPNPs对ANP具有高特异性，而对BSA或混乱ANP肽的亲和力很小。在加入ANP和人血浆的细胞培养基中，MIPNPs也能识别和吸附ANP。综上所述，这些结果表明，MIPNPs对ANP具有高亲和力和选择性，可以作为调节癌症中ANP- npra信号传导的合成抗体。
Natriuretic peptide receptor A (NPRA), the receptor for the cardiac hormone atrial natriuretic peptide (ANP), is expressed abundantly on cancer cells and disruption of ANP-NPRA signaling inhibits tumor burden and metastasis. Since antagonists of NPRA signaling have not provided reproducible results, we reasoned that a synthetic neutralizing antibody to ANP, which has high selectivity and affinity for ANP, could be used to regulate ANP levels and attenuate NPRA signaling. In this study, we prepared molecularly imprinted polymer nanoparticles (MIPNPs) for ANP using a short peptide of ANP as the template and determined their binding affinity and selectivity. The MIPNPs were prepared by precipitation polymerization using NH2-SLRRSS-CONH2, which is a short peptide from ANP as template, methacrylic acid (MAA) and N-isopropylacrylamide (NIPAm) as functional monomers, bis-acrylamide (BIS) as crosslinker. The average diameter of MIPNPs and non-imprinted nanoparticles (NIPNPs) in water is 215.8 ±4.6 nm and 197.7±3.1 nm respectively. The binding isotherm analysis showed that MIPNPs have a much higher binding affinity for template peptide and ANP than NIPNPs. Scatchard analysis gave an equilibrium dissociation constant, Kd of 7.3 μM with a binding capacity 106.7 μmol/g for template peptide and Kd of 7.9 μM with a binding capacity of 36.0 μmol/g for ANP. Measurements of binding kinetics revealed that MIPNPs reach protein adsorption equilibrium in 30 min. MIPNPs found to have high specificity for ANP with little affinity for BSA or scrambled ANP peptide. MIPNPs also recognized and adsorbed ANP in cell culture media spiked with ANP and human plasma. Taken together, these results indicate that MIPNPs have high affinity and selectivity for ANP and can be used as a synthetic antibody for modulating ANP-NPRA signaling in cancers.
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