Cooperative course of Nano-Medical Device Engineering, Hallym University, Chuncheon 24252, Gangwon-do, Korea.
Department of Materials Science and Engineering, Yonsei University, Seoul 03722, Korea.
Sensors (Basel). 2018 Nov 19;18(11):4030. doi: 10.3390/s18114030.
Through orientation control of antibodies, Z-domains autodisplaying outer cell membrane (OM) may be utilized to improve the sensitivity and limit of detection (LOD) of immunoassays and immunosensors. A regenerative immunoaffinity layer based on Z-domains autodisplaying OM was developed for the surface plasmon resonance (SPR) biosensor. Regeneration conditions for the Z-domains autodisplaying OM-based immunoassays and immunosensors were optimized by varying pH and detergent concentration. An cell-based HRP immunoassay was tested and validated in three sequential regenerative immunoassays under optimal conditions. The OM of Z-domains autodisplaying was isolated and coated on the two-dimensional substrate (microplate). The OM-based HRP immunoassay was tested and validated in four regenerative immunoassays. This regenerative OM layer was applied to the SPR biosensor. Z-domains autodisplaying OM layered onto the gold surface of SPR biosensors was developed, and the OM-based regenerative immunoaffinity layer with orientation control was tested using CRP analyte. The SPR biosensor regenerative immunoaffinity layer demonstrated that CRP biosensing was repeated for five regeneration cycles with less than 2% signal difference. Therefore, the newly developed regenerative immunoaffinity layer with antibody orientation control may improve biosensing sensitivity and reduce the cost of medical diagnosis.
通过抗体的定向控制，可以利用自展示外细胞膜（OM）的 Z 结构域来提高免疫分析和免疫传感器的灵敏度和检测限（LOD）。基于自展示 OM 的 Z 结构域开发了一种用于表面等离子体共振（SPR）生物传感器的再生免疫亲和力层。通过改变 pH 值和去污剂浓度优化了基于 Z 结构域自展示 OM 的免疫分析和免疫传感器的再生条件。在最佳条件下，在三个连续的再生免疫分析中测试和验证了基于细胞的 HRP 免疫分析。分离和涂覆有 Z 结构域自展示 OM 的二维基底（微孔板）。在四个再生免疫分析中测试和验证了基于 OM 的 HRP 免疫分析。这种再生 OM 层被应用于 SPR 生物传感器。在 SPR 生物传感器的金表面上开发了自展示 OM 的 Z 结构域，并用 CRP 分析物测试和验证了具有定向控制的 OM 再生免疫亲和力层。SPR 生物传感器的再生免疫亲和力层表明，CRP 生物传感可以重复进行五个再生循环，信号差异小于 2%。因此，具有抗体定向控制的新型再生免疫亲和力层可以提高生物传感的灵敏度并降低医疗诊断的成本。
