RESUMO
Biosensors based on phage display-derived peptides as biorecognition molecules were used for the detection of cell surface cross-species markers in tissue homogenates. The peptide selected for murine myofibers was immobilized onto the surface of an acoustic wave sensor by biotin-streptavidin coupling. To detect peptide-receptor interaction, the sensors were exposed to muscle and control (kidney, liver, brain) tissue homogenates. The sensor showed a strong response to murine muscle. The amplitudes of the responses to the feline muscle homogenates were lower compared to those of the murine muscle, while the same K(d) indicated that the peptide has cross-species affinity. In contrast, murine kidney, liver and brain homogenates produced insignificant responses. Specificity of the sensor was shown in a blocking experiment, as reduced signal was detected when muscle preparations were preincubated with free peptide. Additionally, when muscle-specific peptide was replaced with two different random control peptides, the sensors produced no response to murine muscle. Suitability of peptide ligands for a variety of species can be evaluated using this technology.
Assuntos
Músculo Esquelético/metabolismo , Acústica/instrumentação , Animais , Sítios de Ligação , Técnicas Biossensoriais , Encéfalo/metabolismo , Calibragem , Gatos , Técnicas de Química Combinatória , Técnicas In Vitro , Rim/metabolismo , Fígado/metabolismo , Camundongos , Biblioteca de Peptídeos , Peptídeos/análise , Peptídeos/genética , Peptídeos/metabolismo , Ácidos Esteáricos/metabolismoRESUMO
ASSLNIA, a peptide selected for murine myofibers using phage display technology, was immobilized onto an acoustic wave sensor. The sensor responded to murine and feline muscle homogenates indicating crosspieces interactions. Kidney, liver, and brain preparations produced insignificant responses.