RESUMO
A crystalline silicon surface can be made biocompatible and chemically stable by a self-assembled biofilm of proteins, the hydrophobins (HFBs) purified from the fungus Pleurotus ostreatus. The protein-modified silicon surface shows an improvement in wettability and is suitable for immobilization of other proteins. Two different proteins were successfully immobilized on the HFBs-coated chips: the bovine serum albumin and an enzyme, a laccase, which retains its catalytic activity even when bound on the chip. Variable-angle spectroscopic ellipsometry (VASE), water contact angle (WCA), and fluorescence measurements demonstrated that the proposed approach in silicon surface bioactivation is a feasible strategy for the fabrication of a new class of hybrid devices.
Assuntos
Proteínas Fúngicas/química , Pleurotus/metabolismo , Silício/química , Biocatálise , Biofilmes , Interações Hidrofóbicas e Hidrofílicas , Lacase/metabolismo , Refratometria/métodos , Soroalbumina Bovina/metabolismo , Tensão Superficial , Água/química , MolhabilidadeRESUMO
We have chemically modified the frustules of the marine diatom Coscinodiscus concinnus Wm. Smith to properly bind a highly selective bioprobe such as an antibody. By measuring the changes in the photoluminescence emission of diatoms frustules, we have monitored the molecular recognition event between the antibody and its ligand: the dissociation constant estimated is of the same order of that measured by standard Biacore. The nanostructured silica frustules, a low-cost and natural available material, have shown high sensitivity, equal to 1.2+/-0.2 nm microM(-1), and a detection limit of 100 nM, and thus are quite ideal candidates for lab-on-particle applications.
Assuntos
Técnicas Biossensoriais/instrumentação , Diatomáceas/química , Diatomáceas/ultraestrutura , Imunoensaio/instrumentação , Medições Luminescentes/instrumentação , Desenho de Equipamento , Análise de Falha de Equipamento , Medições Luminescentes/métodos , Biologia Marinha , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Biosilicified nanostructured microshells from the marine diatom Coscinodiscus wailesii have been properly functionalised to bind a molecular probe which specifically recognises a target analyte. The chemical modification process has been characterised by Fourier transformed infrared spectroscopy. Fluorescence measurements demonstrate that the antibodies we used, even if linked to the amorphous silica surface of C. wailesii microshells, still efficiently recognise their antigens. These low cost and largely available natural materials can be thus used as transducers elements for optical biosensors or as targeting microcapsules for drug delivery.