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Advances and Applications in Bioinformatics and Chemistry : AABC (1)
Clinical and Diagnostic Laboratory Immunology (1)
Sensors (Basel, Switzerland) (1)
Fraziano, Maurizio (3)
Colizzi, Vittorio (2)
Amicosante, Massimo (1)
Ausiello, Gabriele (1)
Bianchi, Luigi (1)
Bisen, Prakash S. (1)
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Chandra, Ramesh (1)
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Garg, Sanjay K. (1)
Gherardini, Pier Federico (1)
Ghosh, P. K. (1)
Giacò, Luciano (1)
Helmer-Citterich, Manuela (1)
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In Vitro Analysis of Pyrogenicity and Cytotoxicity Profiles of Flex Sensors to be Used to Sense Human Joint Postures
Santucci, Marilina B.
Sensors (Basel, Switzerland)
Flex sensors can be usefully adopted as mechanical-electrical transducers to measure human joint movements, since their electrical resistance varies proportionally to the angle assumed by the joint under measure. Over time, these sensors have been investigated in terms of mechanical and electrical behavior, but no reports have detailed the possibility of their adoption not just on top but under the human skin of the joint. To this aim, our work investigated in vitro the pyrogenic potential and cytotoxicity of some commercially available flex sensors as a first step toward the necessary requirements regarding their biocompatibility, to predict possible foreign body reactions when used in vivo. Results demonstrated that some specific flex sensors satisfy such requirements.
flex sensors; pyrogenic potential; cytotoxicity; biocompatibility; human joints
B-Pred, a structure based B-cell epitopes prediction server
Gherardini, Pier Federico
Advances and Applications in Bioinformatics and Chemistry : AABC
The ability to predict immunogenic regions in selected proteins by in-silico methods has broad implications, such as allowing a quick selection of potential reagents to be used as diagnostics, vaccines, immunotherapeutics, or research tools in several branches of biological and biotechnological research. However, the prediction of antibody target sites in proteins using computational methodologies has proven to be a highly challenging task, which is likely due to the somewhat elusive nature of B-cell epitopes. This paper proposes a web-based platform for scoring potential immunological reagents based on the structures or 3D models of the proteins of interest. The method scores a protein’s peptides set, which is derived from a sliding window, based on the average solvent exposure, with a filter on the average local model quality for each peptide. The platform was validated on a custom-assembled database of 1336 experimentally determined epitopes from 106 proteins for which a reliable 3D model could be obtained through standard modeling techniques. Despite showing poor sensitivity, this method can achieve a specificity of 0.70 and a positive predictive value of 0.29 by combining these two simple parameters. These values are slightly higher than those obtained with other established sequence-based or structure-based methods that have been evaluated using the same epitopes dataset. This method is implemented in a web server called B-Pred, which is accessible at http://immuno.bio.uniroma2.it/bpred. The server contains a number of original features that allow users to perform personalized reagent searches by manipulating the sliding window’s width and sliding step, changing the exposure and model quality thresholds, and running sequential queries with different parameters. The B-Pred server should assist experimentalists in the rational selection of epitope antigens for a wide range of applications.
B-cell epitopes; immunoinformatics; bioinformatics; web server; epitope prediction
Analysis of the Shotgun Expression Library of the Mycobacterium tuberculosis Genome for Immunodominant Polypeptides: Potential Use in Serodiagnosis
Bisen, Prakash S.
Garg, Sanjay K.
Tiwari, Ram P.
Tagore, P. Ravindra Nath
Ghosh, P. K.
Clinical and Diagnostic Laboratory Immunology
A recombinant DNA strategy was applied to analyze and screen the shotgun expression library from a clinically confirmed local virulent isolate of Mycobacterium tuberculosis with sera from tuberculosis patients, which led to expression and purification of highly immunoreactive and specific mycobacterial antigens expressed during the course of active disease which could be of diagnostic significance. An enzyme-linked immunoassay for diagnosis of tuberculosis was devised by using a shotgun immunoexpression library in the λgt11 vector. DNA from a virulent M. tuberculosis patient isolate (TBW-33) confirmed with the BACTEC 460 system was sheared and expressed to generate shotgun polypeptides. β-Galactosidase fusion proteins capable of demarcating active tuberculosis infections from Mycobacterium bovis BCG-vaccinated healthy subjects or people harboring environmental mycobacteria were selected by comparative immunoreactivity studies. Promising mycobacterial DNA cassettes were subcloned and expressed into the glutathione S-transferase (GST) fusion vector pGEX-5X-1 with a strong tac promoter and were expressed in Escherichia coli BL21. These fusion proteins were severed at a built-in factor Xa recognition site to separate the GST tags and were utilized in an indirect enzyme-linked immunoassay for serodiagnosis of patients with active tuberculosis. The system offered a clear demarcation between BCG-vaccinated healthy subjects and patients with active tuberculosis and proved to be effective in detecting pulmonary as well as extrapulmonary tuberculosis, with an overall sensitivity of 84.33% and an overall specificity of 93.62%.
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