Pilolli Rosa, Visconti Angelo, Monaci Linda
Institute of Sciences of Food Production, National Research Council of Italy (ISPA-CNR), Via Amendola 122/O, 70126, Bari, Italy,
Anal Bioanal Chem. 2015 May;407(13):3787-97. doi: 10.1007/s00216-015-8607-4. Epub 2015 Mar 22.
The development of a surface plasmon resonance (SPR)-based biosensor tailored to the fast detection of egg-related fining allergens in wines is herein described. Ovalbumin (OVA) was chosen as the target protein to be monitored due to its highest abundance in the egg white powder, a typical fining agent used by the winery industry to promote wine clarification. A direct assay was designed, basing on the use of polyclonal anti-OVA antibody as bio-specific receptor. With the aim of optimizing the assay conditions, different parameters able to influence the final biosensor response were carefully investigated (i.e., pH, ionic strength, and additional surfactant concentration). After the fine tuning of these parameters, the assay was tested in the direct analysis of OVA in commercial wines artificially contaminated with egg white powder at different concentration levels in order to assess the reliability of the biosensor in detecting traces of OVA in complex matrices. The devised assay allowed to trace, in a short analysis time and with a minimal sample pre-treatment required, the presence of egg allergens at the lowest concentration comprised between 0.03 and 0.2 μg/mL. Finally, the response provided by the developed biosensor was correlated with an established liquid chromatography mass spectrometry (LC-MS) method developed in our laboratories, and performances of both approaches were assessed for the fast monitoring of egg allergen contamination in fined wines.
本文描述了一种基于表面等离子体共振(SPR)的生物传感器的开发,该传感器专为快速检测葡萄酒中与蛋清相关的澄清过敏原而设计。由于卵清蛋白(OVA)在蛋白粉中含量最高,而蛋白粉是酿酒行业用于促进葡萄酒澄清的一种典型澄清剂,因此选择OVA作为要监测的目标蛋白。设计了一种直接检测方法,基于使用多克隆抗OVA抗体作为生物特异性受体。为了优化检测条件,仔细研究了能够影响最终生物传感器响应的不同参数(即pH值、离子强度和额外的表面活性剂浓度)。在对这些参数进行微调后,该检测方法用于直接分析人工添加不同浓度蛋白粉的市售葡萄酒中的OVA,以评估生物传感器在复杂基质中检测痕量OVA的可靠性。所设计的检测方法能够在短时间内进行分析,且所需样品预处理最少,检测出最低浓度在0.03至0.2μg/mL之间的蛋清过敏原。最后,将所开发生物传感器的响应与我们实验室建立的液相色谱质谱(LC-MS)方法进行关联,并评估两种方法在快速监测澄清葡萄酒中蛋清过敏原污染方面的性能。