Amino acid analysis of lipases from oil pollutant isolates: Cunninghamella verticillata and Geotrichum candidum

Lipase is an enzyme commonly used in food, dairy, and other industries. Fungal lipases are more commonly used due to their secretion and easier production. Analyses of the amino acid composition of microbial lipases will hasten their potential usage in industrial applications. In this study, the major amino acid compositions of lipases secreted by oil pollutant isolates (Cunninghamella verticillata and Geotrichum candidum) enriched with fatty substances were analyzed by high performance liquid chromatography. Among eight major amino acids found in these lipases, histidine and ornithine were predominant. Lysine was absent from lipase generated by C. verticillata, while glutamine was absent from that produced by G. candidum. Conversely, glutamic acid, asparagine, histidine and arginine were present in slightly higher amounts in G. candidum. However, a slight decrease in aspartic acid and ornithine was observed in G. candidum. Analyses of the amino acids composition of lipase can potentially facilitate to predict the nature of this enzyme

Voltammetric aptasensors for protein disease biomarkers detection: a review

"Available online 24 May 2016"An electrochemical aptasensor is a compact analytical device where the bioreceptor (aptamer) is coupled to a transducer surface to convert a biological interaction into a measurable signal (current) that can be easily processed, recorded and displayed. Since the discovery of the Systematic Evolution of Ligands by Enrichment (SELEX) methodology, the selection of aptamers and their application as bioreceptors has become a promising tool in the design of electrochemical aptasensors. Aptamers present several advantages that highlight their usefulness as bioreceptors such as chemical stability, cost effectiveness and ease of modification towards detection and immobilization at different transducer surfaces. In this review, a special emphasis is given to the potential use of electrochemical aptasensors for the detection of protein disease biomarkers using voltammetry techniques. Methods for the immobilization of aptamers onto electrode surfaces are discussed, as well as different electrochemical strategies that can be used for the design of aptasensors.The authors acknowledge the financial support from the Strategic funding of UID/BIO/04469/2013 unit, from Project POCI-01-0145- FEDER-006984 – Associate Laboratory LSRE-LCM funded by FEDER funds through COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI) – and by national funds through FCT - Fundação para a Ciência e a Tecnologia and project ref. RECI/BBB-EBI/ 0179/2012 (project number FCOMP-01-0124-FEDER-027462) and S. Meirinhos's doctoral grant (ref SFRH/BD/65021/2009) funded by Fundação para a Ciência e a Tecnologia

Aptamers as a replacement for antibodies in enzyme-linked immunosorbent assay

The application of antibodies in enzyme-linked immunosorbent assay (ELISA) is the basis of this diagnostic technique which is designed to detect a potpourri of complex target molecules such as cell surface antigens, allergens, and food contaminants. However, development of the systematic evolution of Ligands by Exponential Enrichment (SELEX) method, which can generate a nucleic acid-based probe (aptamer) that possess numerous advantages compared to antibodies, offers the possibility of using aptamers as an alternative molecular recognition element in ELISA. Compared to antibodies, aptamers are smaller in size, can be easily modified, are cheaper to produce, and can be generated against a wide array of target molecules. The application of aptamers in ELISA gives rise to an ELISA-derived assay called enzyme-linked apta-sorbent assay (ELASA). As with the ELISA method, ELASA can be used in several different configurations, including direct, indirect, and sandwich assays. This review provides an overview of the strategies involved in aptamer-based ELASA. (C) 2014 Elsevier B.V. All rights reserved

Aptamers as a replacement for antibodies in enzyme-linked immunosorbent assay

The application of antibodies in enzyme-linked immunosorbent assay (ELISA) is the basis of this diagnostic technique which is designed to detect a potpourri of complex target molecules such as cell surface antigens, allergens, and food contaminants. However, development of the systematic evolution of Ligands by Exponential Enrichment (SELEX) method, which can generate a nucleic acid-based probe (aptamer) that possess numerous advantages compared to antibodies, offers the possibility of using aptamers as an alternative molecular recognition element in ELISA. Compared to antibodies, aptamers are smaller in size, can be easily modified, are cheaper to produce, and can be generated against a wide array of target molecules. The application of aptamers in ELISA gives rise to an ELISA-derived assay called enzyme-linked apta-sorbent assay (ELASA). As with the ELISA method, ELASA can be used in several different configurations, including direct, indirect, and sandwich assays. This review provides an overview of the strategies involved in aptamer-based ELASA. (C) 2014 Elsevier B.V. All rights reserved

Current aspects in immunosensors

Sensing applications can be used to report biomolecular interactions in order to elucidate the functions of molecules. The use of an analyte and a ligand is a common set-up in sensor development. For several decades, antibodies have been considered to be potential analytes or ligands for development of socalled "immunosensors." In an immunosensor, formation of the complex between antibody and antigen transduces the signal, which is measurable in various ways (e.g., both labeled and label-free based detection). Success of an immunosensor depends on various factors, including surface functionalization, antibody orientation, density of the antibody on the sensor platform, and configuration of the immunosensor. Careful optimization of these factors can generate clear-cut results for any immunosensor. Herein, current aspects, involved in the generated immunosensors, are discussed