Method To Determine Protein Concentration in the Protein Nanoparticle Conjugates Aqueous Solution Using Circular Dichroism Spectroscopy
Method To Determine Protein Concentration in the Protein Nanoparticle Conjugates Aqueous Solution Using Circular Dichroism Spectroscopy
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DOI：
10.1021/acs.analchem.5b01451
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发表时间：
2015-07-07
影响因子：
7.4
通讯作者：
Leblanc, Roger M.
中科院分区：
文献类型：
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作者：
Li, Shanghao;Peng, Zhili;Leblanc, Roger M.
为了合成和表征蛋白质-纳米粒子缀合物（蛋白质-NP），人们付出了巨大的努力，以期它们在生物纳米技术中的应用前景良好。然而，迄今为止，蛋白质-NN中的蛋白质浓度测定尚未见报道。在这封信中，我们提出了一种简单且无损的方法，使用圆二色性 (CD) 光谱来量化蛋白质纳米颗粒水溶液中的蛋白质浓度。将碳点（类似于 4 nm）、金纳米颗粒（类似于 40 nm）和聚乙二醇（PEG，分子量类似于 3000）与蛋白质（人转铁蛋白、人血清白蛋白和卵清蛋白）物理混合或共价结合（金纳米颗粒除外）。我们能够使用圆二色光谱的校准曲线来量化蛋白质-纳米颗粒缀合物中的蛋白质浓度。
Considerable efforts have been made to synthesize and characterize protein-nanopartide conjugates (protein-NPs) for their promising applications in bionanotechnology. However, protein concentration determination in the protein-NN has so far not been reported. In this Letter, we present a simple and nondestructive approach to quantify the protein concentration in the protein-NPs aqueous solution using circular dichroism (CD) spectroscopy. Carbon dots (similar to 4 nm), gold nanopartides (similar to 40 nm), and polyethylene glycol (PEG, molecular weight similar to 3000) were either physically mixed or covalently conjugated (not in the case of gold nanoparticles) with proteins (human transferrin, human serum albumin, and ovalbumin). We were able to quantify the protein concentration in the protein-nanoparticle conjugates using a calibration curve from the CD spectra.