Molecularly Imprinted SPR Sensor for Detection of β-Lactoglobulin in Camel Milk Powder
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摘要: 结合模板分子前定位、表面印迹法和后修饰策略，在金（Au）基底上构建了分子印迹表面等离子体共振（SPR）传感器，并将其用于骆驼奶粉中β-乳球蛋白（BLG）的检测。首先，在聚多巴胺（PDA）修饰的金片（PDA/Au）上，通过聚丙烯酸（PAA）固定模板分子BLG，以多巴胺为功能单体和交联剂进行印迹，然后在非印迹部分接枝具有抗蛋白质吸附功能的部分水解的聚（2-甲基-2-噁唑啉）（PMOXA-EI），洗脱模板后，得到BLG分子印迹SPR传感器。采用水接触角（WCA）研究了传感器表面亲/疏水的变化，用傅里叶变换红外光谱仪（FT-IR）、原子力显微镜（AFM）、可变角光谱椭偏仪（VASE）对其进行表征。结果表明，该传感器对0.21 ～ 10 μg/mL的BLG具有良好的线性关系，检测限为0.12 μg/mL，能快速灵敏地检测出骆驼奶粉中BLG的质量浓度，回收率在100.2% ～ 100.7%。
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关键词:
- β-乳球蛋白 /
- 多巴胺 /
- 分子印迹 /
- SPR传感器 /
- 聚(2-甲基-2-噁唑啉)
Abstract: Combined with template molecular pre-positioning, surface imprinting method, and post-modification strategy, a molecularly imprinted surface plasmon resonance (SPR) sensor was constructed on a gold substrate and used for detection of β-lactoglobulin (BLG) in camel milk powder. Firstly, the template molecule BLG was fixed by polyacrylic acid (PAA) on the gold substrate which was modified by polydopamine in advance. Then imprinting was performed using dopamine as functional monomer and crosslinking agent. Finally, partially hydrolyzed poly (2-methyl-2-oxazoline) (PMOXA-EI) with anti-protein adsorption function was grafted on the non-imprinted part, and the BLG molecularly imprinted SPR sensor was prepared after eluting the template. The changes of sensor’s surface hydrophilicity/hydrophobicity were studied by water contact angle (WCA) and the characterization of created sensor was performed by using Fourier transform infrared spectrometer (FT-IR), atomic force microscopy (AFM), and variable angle spectroscopy ellipsometry (VASE), respectively. Results show that the sensor performe a good linear relationship in the range of 0.21～10 μg/mL for BLG, and the detection of limit is 0.12 μg/mL. The sensor could detect the BLG in camel milk powder quickly and sensitively with a recovery rate between 100.2% and 100.7%.-
Key words:
- β-lactoglobulin /
- dopamine /
- molecular imprinting /
- SPR sensor /
- poly(2-methyl-2-oxazoline)
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图 1 MIP SPR传感器的制备示意图
Figure 1. Schematic illustration for the preparation of MIP SPR sensor
图 2 MIP-before elution、MIP和NIP的红外光谱
Figure 2. FT-IR spectra of MIP-before elution, MIP and NIP
图 5 BLG在MIP传感器上的（a）等温吸附线和（b）标准校准曲线
Figure 5. (a) Adsorption isotherms and (b) standard calibration curve of BLG on the MIP sensor
图 6 MIP传感器对BLG吸附的重复性和再现性
Figure 6. Repeatability and reproducibility of BLG adsorption by MIP sensor
表 1 生物传感器对BLG和竞争蛋白的Q、IF和k
Table 1. Q、IF and k of BLG and competing proteins by biosensor
Protein QMIP/RU QNIP/RU IF k BLG 1401 458 3.1 — SA 647 690 0.9 2.2 LYZ 254 363 0.7 5.5 ALB 527 574 0.9 2.6 LF 203 218 0.9 6.9 
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表 2 骆驼奶粉中BLG质量浓度的检测
Table 2. Detection of BLG mass concentration in camel milk powder
ρ（Spiked BLG）/（μg·mL−1） ρ（Found BLG）/（μg·mL−1） Recovery/% 3.33 3.35 ± 0.25 100.6 ± 7.5 5.00 5.01 ± 0.15 100.2 ± 3.0 10.00 10.07 ± 0.14 100.7 ± 1.4
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