Use of an immunoenzyme method for detecting specific antibodies to Pseudomonas aeruginosa in patients with acute and chronic lung diseases

ELISA in which P. aeruginosa slime preparations are used as antigens permits the detection of antibodies to this microorganism in the blood sera of patients with acute and chronic pulmonary diseases induced by P. aeruginosa. This assay makes it possible to find out the etiology of infection even before the results of bacteriological investigation are obtained.     

A versatile tool in controlling aggregation and Ag nanoparticles assisted in vitro folding of thermally denatured zDHFR

BackgroundProteins have tendency to form inactive aggregates at higher temperatures due to thermal instability. Maintenance of thermal stability is essential to gain the protein in sufficient quantity and biologically active form during their commercial production.MethodsBL21-DE3 Rosetta E. coli cells which contains plasmid pET43.1a vector was used for producing zDHFR protein commercially. The purification of N-terminal Histidine tagged zDHFR was performed by Immobilized Metal Ion chromatography (IMAC). Investigations were performed in existence and non existence of Silver nanoparticles (AgNPs). The inactivation kinetics of zDHFR in existence and non existence of AgNPs were monitored over a range of 40–80 °C as monitored by UV–Visible absorption spectroscopy.ResultsThe protein completely lost its activity at 55 °C. Kinetics of inactivated zDHFR follows first order model in presence and absence of AgNPs. Decrease in rate constant (k) values at respective temperatures depicts that AgNPs contribute in the thermostability of the protein. AgNPs also assists in regaining the activity of zDHFR protein.ConclusionsAgNPs helps in maintaining thermostability and reducing the aggregation propensity of zDHFR protein.General significanceResult explains that AgNPs are recommended as a valuable system in enhancing the industrial production of biologically active zDHFR protein which is an important component in folate cycle and essential for survival of cells and prevents the protein from being aggregated.