Frozen assembly of gold nanoparticles for rapid analysis of antifreeze protein activity.
We report the novel activity-based detection of antifreeze protein (AFP), also known as ice-binding protein (IBP), using freeze-labile gold nanoparticles (AuNPs) in order to overcome labor-intensive and low throughput issues of the current method based on thermal hysteresis (TH). Upon the addition of either CnAFP from the Antarctic diatom Chaetoceros neogracile or LeIBP from the Arctic yeast Leucosporidium sp. to mercaptosuccinic acid-capped AuNP, the self-assembly of AuNPs was highly inhibited after a freezing/thawing cycle, leading to no color change in the AuNP solution. As a result, the aggregation parameter (E(520)/E(650)) of AuNP presented the rapid detection of both the concentration-dependent activity and stability of two AFPs with high sensitivity, where the detection range was 100-fold lower than that of the TH-based method. We suggest that our newly developed method is very suitable for simple and high-throughput measurement of AFP activity.
Department of Life Science, Hanyang University, Seoul 133-791, Republic of Korea.
Ji-In
JI
Park
Jun Hyuck
JH
Lee
Yunho
Y
Gwak
Hak Jun
HJ
Kim
EonSeon
E
Jin
Young-Pil
YP
Kim
D021301:Antifreeze Proteins / Q000032:analysis* / Q000737:chemistry*; D015374:Biosensing Techniques / Q000295:instrumentation*; D003124:Colorimetry / Q000295:instrumentation*; D003460:Crystallization / Q000379:methods*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005615:Freezing; D006046:Gold / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D021301:Antifreeze Proteins; D006046:Gold