Ribozyme-mediated signal augmentation on a mass-sensitive biosensor.
Mass-based detection methods such as the quartz crystal microbalance (QCM) offer an attractive option to label-based methods; however the sensitivity is generally lower by comparison. In particular, low-molecular-weight analytes can be difficult to detect based on mass addition alone. In this communication, we present the use of effector-dependent ribozymes (aptazymes) as reagents for augmenting small ligand detection on a mass-sensitive device. Two distinct aptazymes were chosen: an L1-ligase-based aptazyme (L1-Rev), which is activated by a small peptide (MW approximately 2.4 kDa) from the HIV-1 Rev protein, and a hammerhead cleavase-based aptazyme (HH-theo3) activated by theophylline (MW = 180 Da). Aptazyme activity was observed in real time, and low-molecular-weight analyte detection has been successfully demonstrated with both aptazymes.
Knudsen
Scott M
SM
Department of Chemistry and Biochemistry, University of Texas at Austin, Austin, Texas 78712, USA.
Lee
Joonhyung
J
Ellington
Andrew D
AD
Savran
Cagri A
CA
D015374:Biosensing Techniques / Q000379:methods*; D008025:Ligases / Q000378:metabolism; D008970:Molecular Weight; D009690:Nucleic Acid Conformation; D012313:RNA / Q000737:chemistry; D016337:RNA, Catalytic / Q000737:chemistry / Q000378:metabolism*
D016337:RNA, Catalytic; D012313:RNA; D008025:Ligases