title: Reciprocating-flow ATP amplification system for increasing the number of amplification cycles.
abstract: 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.
authors.affiliation: Research Institute for Nanodevice and Bio Systems, Hiroshima University, Higashi-Hiroshima, Hiroshima 739-8530, Japan.
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Satoh
authors.forename: Kosuke
authors.initials: K
authors.lastname: Tsuruta
authors.forename: Yasuharu
authors.initials: Y
authors.lastname: Shinoda
authors.forename: Ryuichi
authors.initials: R
authors.lastname: Hirota
authors.forename: Kenichi
authors.initials: K
authors.lastname: Noda
authors.forename: Akio
authors.initials: A
authors.lastname: Kuroda
authors.forename: Yuji
authors.initials: Y
authors.lastname: Murakami
mesh_terms: D000255:Adenosine Triphosphate / Q000032:analysis* / Q000378:metabolism; D000263:Adenylate Kinase / Q000378:metabolism; D055767:Immobilized Proteins / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D011770:Pyruvate Kinase / Q000378:metabolism
chemical_list: D055767:Immobilized Proteins; D000255:Adenosine Triphosphate; D011770:Pyruvate Kinase; D000263:Adenylate Kinase