A continuous-flow ATP amplification system for increasing the sensitivity of quantitative bioluminescence assay

Satoh Tetsuya

Shinoda Yasuharu

Alexandrov Maxym

Kuroda Akio

Murakami Yuji

Analytical Biochemistry

We constructed a novel ATP amplification reactor using a continuous-flow system, and this allowed us to increase the sensitivity of quantitative bioluminescence assay by controlling the number of ATP amplification cycles. We previously developed a bioluminescence assay coupled with ATP amplification using a batch system. However, it was difficult to control the number of amplification cycles. In this study, ATP amplification was performed using a continuous-flow system, and significant linear correlations between amplified luminescence and initial ATP concentration were observed. When performing 4 cycles of continuous-flow ATP amplification, the gradient of amplification was 1.87N. Whereas lower quantifiable level was 500 pM without amplification, values as low as 50 pM ATP could be measured after amplification. Sensitivity thus increased 10-fold, with further improvements expected with additional amplification cycles. The continuous-flow system thus effectively increased the sensitivity of quantitative bioluminescence assay.

Bioluminescence assay

ATP amplification

Continuous-flow system

Biology [ 460 ]

eng

Academic Press Inc.

Elsevier Science

Copyright (c) 2008 Elsevier Inc.

[ISSN] 0003-2697

[DOI] 10.1016/j.ab.2008.04.045

[NCID] AA00524867

[DOI] http://dx.doi.org/10.1016/j.ab.2008.04.045