School of Chemical Engineering, Sungkyunkwan University, 300 Cheoncheon-dong, Jangan-gu, Suwon, Gyeonggi-do 440-746, South Korea.
Biosens Bioelectron. 2010 Dec 15;26(4):1548-53. doi: 10.1016/j.bios.2010.07.109. Epub 2010 Aug 4.
In this study, hybrid stimulus was initially introduced to improve the sensitivity of PDA vesicle chip for detection of prostate-specific antigen-α1-antichymotrypsin (PSA-ACT) complex. The strategy of hybrid stimulus on PDA vesicle chip offers the amplification method of fluorescent signal which combines a primary response by the immune reaction of antigen-antibody and a secondary response by the mechanical pressure of pAb-conjugated magnetic beads. As the primary response result on PDA vesicle chip, the PSA-ACT complex in PBS buffer was detected at 10 ng/mL. However, this detection sensitivity was insufficient for diagnosis of prostate cancer because the normal human PSA concentration is less than 4.0 ng/mL. To solve this problem, polyclonal PSA antibody-conjugated magnetic beads were used as an amplifying agent after primary immunoresponse. As a result, the PSA-ACT complex concentrations (as low as 0.1 ng/mL) could be detected in the PBS buffer sample. Therefore, this result can be applied to various fields, such as the detection of cells, proteins, and DNA for sensitive and specific biosensing based on PDA supramolecules.
在这项研究中，我们最初引入混合刺激来提高 PDA 囊泡芯片检测前列腺特异性抗原-α1-抗糜蛋白酶 (PSA-ACT) 复合物的灵敏度。PDA 囊泡芯片上的混合刺激策略提供了荧光信号放大方法，该方法结合了抗原-抗体免疫反应的初级反应和 pAb 偶联磁珠的机械压力的次级反应。作为 PDA 囊泡芯片上的初级反应结果，在 PBS 缓冲液中检测到 10ng/mL 的 PSA-ACT 复合物。然而，由于正常人 PSA 浓度低于 4.0ng/mL，因此这种检测灵敏度不足以用于诊断前列腺癌。为了解决这个问题，在初级免疫反应后，使用多克隆 PSA 抗体偶联磁珠作为放大剂。结果，在 PBS 缓冲液样品中可以检测到浓度低至 0.1ng/mL 的 PSA-ACT 复合物。因此，该结果可应用于各种领域，例如细胞、蛋白质和 DNA 的检测，用于基于 PDA 超分子的敏感和特异性生物传感。
