Reciprocating-flow ATP amplification system for increasing the number of amplification cycles

Satoh Tetsuya

Tsuruta Kosuke

Shinoda Yasuharu

Hirota Ryuichi

Noda Kenichi

Kuroda Akio

Murakami Yuji

Analytical Biochemistry

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 ATIP 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 oil the columns was 0.03 MPa in 1-10 ATIP amplification cycles, and no increases in backpressure were observed.

ATP amplification

Reciprocating-flow system

Bioluminescence assay

Biology [ 460 ]

eng

Academic Press INC

Elsevier Science

Copyright (c) 2009 Elsevier Inc.

[ISSN] 0003-2697

[DOI] 10.1016/j.ab.2009.08.025

[NCID] AA00524867

[DOI] http://dx.doi.org/10.1016/j.ab.2009.08.025