Surface plasmon resonance biosensing: Approaches for screening and characterising antibodies for food diagnostics.
Surface plasmon resonance biosensing: Approaches for screening and characterising antibodies for food diagnostics.
复制标题
DOI：
10.1016/j.talanta.2016.05.008
复制
发表时间：
2016-08-15
期刊：
影响因子：
6.1
通讯作者：
Campbell K
中科院分区：
文献类型：
--
作者：
Yakes BJ;Buijs J;Elliott CT;Campbell K
过去二十年来，作为食品诊断替代技术来检测化学污染物和食源性病原体的生物传感方法的研究有所增加。此类测试的关键组成部分是生物识别元件，多克隆或单克隆抗体仍然在市场上占据主导地位。传统上，分别通过酶免疫测定来筛选血清或细胞培养基以选择多克隆或单克隆候选抗体。对于利基毒素化合物，由于用于缀合物生产的毒素供应的限制，酶免疫测定对于抗体生产来说可能是昂贵的和/或令人望而却步的方法。在食品诊断中经过验证的自动化、自我再生、基于芯片的生物传感器可用作候选抗体选择的快速筛选工具。这项工作描述了使用单通道和多通道表面等离子共振 (SPR) 生物传感器来选择和表征抗体，以及它们在贝类组织中的评估，作为检测软骨藻酸（作为模型毒素化合物）的标准技术。与酶免疫测定相比，使用这些生物传感器技术筛选单克隆抗体生产中的杂交瘤的主要优点是实时观察分子相互作用和快速的分析周转时间。多通道原型仪器在 2 小时内完成了 96 次分析，而单通道需要 12 小时，而 ELISA 免疫测定则需要超过 24 小时。选择高灵敏度抗体，IC50 范围为单克隆 4.8 至 6.9 ng/mL，多克隆为 2.3–6.0 ng/mL，用于在 1 分钟分析时间内检测软骨藻酸。尽管生物传感器技术在食品工业的低成本、多重便携式诊断方面取得了进展，但如本文所示，用于食品诊断抗体开发的基于实验室的 SPR 仪器仍然占有一席之地。
Research in biosensing approaches as alternative techniques for food diagnostics for the detection of chemical contaminants and foodborne pathogens has increased over the last twenty years. The key component of such tests is the biorecognition element whereby polyclonal or monoclonal antibodies still dominate the market. Traditionally the screening of sera or cell culture media for the selection of polyclonal or monoclonal candidate antibodies respectively has been performed by enzyme immunoassays. For niche toxin compounds, enzyme immunoassays can be expensive and/or prohibitive methodologies for antibody production due to limitations in toxin supply for conjugate production. Automated, self-regenerating, chip-based biosensors proven in food diagnostics may be utilised as rapid screening tools for antibody candidate selection. This work describes the use of both single channel and multi-channel surface plasmon resonance (SPR) biosensors for the selection and characterisation of antibodies, and their evaluation in shellfish tissue as standard techniques for the detection of domoic acid, as a model toxin compound. The key advantages in the use of these biosensor techniques for screening hybridomas in monoclonal antibody production were the real time observation of molecular interaction and rapid turnaround time in analysis compared to enzyme immunoassays. The multichannel prototype instrument was superior with 96 analyses completed in 2 h compared to 12 h for the single channel and over 24 h for the ELISA immunoassay. Antibodies of high sensitivity, IC50's ranging from 4.8 to 6.9 ng/mL for monoclonal and 2.3–6.0 ng/mL for polyclonal, for the detection of domoic acid in a 1 min analysis time were selected. Although there is a progression for biosensor technology towards low cost, multiplexed portable diagnostics for the food industry, there remains a place for laboratory-based SPR instrumentation for antibody development for food diagnostics as shown herein.