Research Institute for Nanodevice and Bio Systems, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8530, Japan.
Anal Biochem. 2009 Dec 15;395(2):161-5. doi: 10.1016/j.ab.2009.08.025. Epub 2009 Aug 21.
We constructed a novel ATP amplification reactor using a reciprocating-flow system to increase the number of ATP amplification cycles without an increase in backpressure. We previously reported a continuous-flow ATP amplification system that effectively and quantitatively amplified ATP and increased the sensitivity of a quantitative bioluminescence assay. However, it was difficult to increase the number of amplification cycles due to backpressure in the system. Because addition of immobilized adenylate kinase (ADK) and pyruvate kinase (PK) columns increased backpressure, the maximum number of ATP amplification cycles within column durability was only 4. In this study, ATP amplification was performed using a reciprocating-flow system, and 10 cycles of ATP amplification could be achieved without an increase in backpressure. As a result, ATP was amplified more than 100-fold after 10 cycles of reciprocating flow. The gradient of ATP amplification was approximately 1.76(N). The backpressure on the columns was 0.03 MPa in 1-10 ATP amplification cycles, and no increases in backpressure were observed.
我们构建了一种新型的 ATP 扩增反应器，使用往复流系统在不增加背压的情况下增加 ATP 扩增循环的次数。我们之前报道了一种连续流动的 ATP 扩增系统，该系统可以有效地、定量地扩增 ATP，并提高定量生物发光测定的灵敏度。然而，由于系统中的背压，增加扩增循环的次数变得困难。因为添加固定化腺苷酸激酶（ADK）和丙酮酸激酶（PK）柱会增加背压，所以在柱耐久性内的最大 ATP 扩增循环次数仅为 4 次。在这项研究中，使用往复流系统进行 ATP 扩增，在不增加背压的情况下可以实现 10 次 ATP 扩增循环。结果，经过 10 次往复流循环后，ATP 被扩增了 100 多倍。ATP 扩增的梯度约为 1.76（N）。在 1-10 次 ATP 扩增循环中，柱上的背压为 0.03 MPa，没有观察到背压的增加。
