Development of biosensors with aptamers as bio-recognition element: the case of HIV-1 Tat protein.
The in vitro selection of combinatorial libraries of RNA/DNA, has allowed the identification of specific nucleic acids (aptamers) which bind to a wide range of target molecules with high affinity and specificity. In this work, an RNA aptamer, specific for the protein trans-activator of transcription (Tat) of HIV-1, has been used as bio-recognition element to develop a biosensor (aptasensor). The biosensor was optimised using piezoelectric quartz-crystals as transducers and the aptamer was immobilised on the gold electrode of the crystal. The immobilisation procedure was based on the interaction between the biotinylated aptamer and streptavidin previously deposited on the electrode. The main analytical characteristics of the biosensor, such as sensitivity, selectivity and reproducibility, have been studied in details. An optimised regeneration procedure allowed the multiple use of the aptamer-coated crystal. The aptasensor has been compared with the corresponding immunosensor, based on the specific monoclonal anti-Tat antibody. The antibody was immobilised on a layer of carboxylated dextran previously deposited on the gold electrode. The results demonstrated that the use of a biosensor with a specific aptamer as bio-recognition element could be an interesting approach in the detection of proteins, which has been here examined considering a model system.
Minunni
M
M
Università degli Studi di Firenze, Dipartimento di Chimica, Via della Lastruccia 3, 50019 Sesto Fiorentino (FI), Italy.
Tombelli
S
S
Gullotto
A
A
Luzi
E
E
Mascini
M
M
D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D020099:Coated Materials, Biocompatible / Q000737:chemistry*; D004563:Electrochemistry / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D015696:Gene Products, tat / Q000032:analysis* / Q000737:chemistry*; D015497:HIV-1 / Q000378:metabolism*; D012313:RNA / Q000737:chemistry*; D016601:RNA-Binding Proteins / Q000032:analysis / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D054322:tat Gene Products, Human Immunodeficiency Virus
D020099:Coated Materials, Biocompatible; D015696:Gene Products, tat; D016601:RNA-Binding Proteins; D054322:tat Gene Products, Human Immunodeficiency Virus; D012313:RNA