A cell-based immunobiosensor with engineered molecular recognition .1. Design feasibility
A cell-based immunobiosensor with engineered molecular recognition .1. Design feasibility
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DOI：
10.1016/s0956-5663(96)00068-1
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发表时间：
1997-01-01
影响因子：
12.6
通讯作者：
Page, DL
中科院分区：
文献类型：
--
作者：
Pizziconi, VB;Page, DL
本发明描述了一种新型生物电子传感器，其中活免疫细胞通过工程化其对临床感兴趣的预选抗原的分子识别而转化成独特的生物换能器对。这种"混合"生物传感器，与肥大细胞接口的微制造的热电装置与使用生物分子连接，是能够检测抗原在真实的时间，通过转换从抗原诱导的肥大细胞活化过程中产生的微小的热变化。热电方法的选择基于初步的生物能量计算，这表明肥大细胞抗原识别引起的代谢变化导致相对于基础代谢条件的放热显着增加。实验研究证实，肥大细胞活化和脱粒可以区分从基础代谢活动热。从使用培养的肥大细胞（MC/9粘膜样肥大细胞系）和收获的肥大细胞（大鼠腹膜肥大细胞）的微量热实验获得的结果表明，在细胞活化后立即可检测到热输出的增加（类似于3 +/-0.5pW/细胞，平均峰值输出）。一个微型的混合免疫生物传感器设备的建设是可能的生物电子耦合实现与使用的细胞粘附蛋白，固定非粘附（MC/9）细胞以及粘附（RBL-2H3大鼠嗜碱性白血病）细胞的热电堆。在混合生物传感器原型上进行的初步测试的结果验证了微型活细胞免疫诊断生物传感器的设计可行性。这种基于细胞的混合生物传感器方法可以极大地扩展对广泛的临床相关抗原的选择性、快速、现场抗原检测的能力，并提供体外诊断的新方法。(C)1997年爱思唯尔科学有限公司。
A novel bioelectronic sensor is described in which living immune cells are transformed into unique biotransducer couples by engineering their molecular recognition for preselected antigens of clinical interest. This 'hybrid' biosensor, constructed with mast cells interfaced to a microfabricated thermoelectric device with the use of biomolecular linkages, is capable of detecting antigens in real time by transducing minute heat changes arising from antigen-induced mast cell activation processes. The thermoelectric approach was selected based upon preliminary bioenergetic calculations which indicated that metabolic changes arising from mast cell antigen recognition result in a significant increase in exothermic heat relative to basal metabolic conditions. Experimental studies confirmed that mast cell activation and degranulation can be discriminated thermally from basal metabolic activity. Results obtained from microcalorimetry experiments using cultured mast cells (MC/9 mucosal-like mast cell line), and harvested mast cells (rat peritoneal mast cells) indicated that detectable increases in heat output (similar to 3 +/- 0.5 pW/cell, mean peak output) immediately followed cell activation. The construction of a miniature hybrid immunobiosensor device was made possible by bioelectronic coupling achieved with the use of cellular adhesive proteins that immobilized non-adherent (MC/9) cells as well as adherent (RBL-2H3 rat basophilic leukemia) cells to the thermopile. Results from preliminary tests conducted on a hybrid biosensor prototype validated the design feasibility of a miniature, living cell immunodiagnostic biosensor. Such cell-based hybrid biosensor approaches may greatly extend the capability for selective, rapid, on-site, antigen detection for a wide range of clinically relevant antigens and offer new approaches to in vitro diagnostics. (C) 1997 Elsevier Science Limited.