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Department of Pharmacology

 
Read more at: Design of novel affinity adsorbents for the purification of trypsin‐like proteases

Design of novel affinity adsorbents for the purification of trypsin‐like proteases

A number of ligands for the selective purification by affinity chromatography of the trypsin‐like protease, porcine pancreatic kallikrein, were designed de novo by computer‐aided molecular design. The ligands were designed to mimic the side‐chains of a number of arginyl dipeptides and included a benzamidine moiety substituted on a triazine ring. The ligands displayed inhibitory activities against pancreatic kallikrein which mirrored the specificity constants of the dipeptides they were designed to mimic.


Read more at: Designer dyes: 'biomimetic' ligands for the purification of pharmaceutical proteins by affinity chromatography

Designer dyes: 'biomimetic' ligands for the purification of pharmaceutical proteins by affinity chromatography

Affinity chromatography has been extensively refined over the past few years to meet the more stringent criteria being placed on recombinant proteins as therapeutic products. New developments in the design of selective and stable ligands for affinity chromatography are establishing the technique as a routine tool in process-scale protein purification. Exploitation of sophisticated molecular modelling techniques in conjunction with binding and crystallographic studies has permitted the design of new, highly selective 'biomimetic' ligands for the target proteins. © 1992.


Read more at: Novel affinity separations based on perfluorocarbon emulsions. Development of a perfluorocarbon emulsion reactor for continuous affinity separations and its application in the purification of human serum albumin from blood plasma

Novel affinity separations based on perfluorocarbon emulsions. Development of a perfluorocarbon emulsion reactor for continuous affinity separations and its application in the purification of human serum albumin from blood plasma

Perfluorocarbon affinity emulsions are generated by the homogenisation of a perfluorocarbon oil with a polymeric fluorosurfactant previously derivatised with an affinity ligand and subsequently cross-linked in situ. This procedure gives rise to a novel liquid affinity adsorbent that can be used for continuous protein purification. Discrete emulsion droplets were found to be unstable when pumped for prolonged periods; however, when flocculated, the emulsion floccules with diameters of around 125 μm, were very stable and sedimented faster.


Read more at: Nucleotide sequence and over-expression of morphine dehydrogenase, a plasmid-encoded gene from Pseudomonas putida M10

Nucleotide sequence and over-expression of morphine dehydrogenase, a plasmid-encoded gene from Pseudomonas putida M10

Pseudomonas putida M10 was originally isolated from factory waste liquors by selection for growth on morphine. The NADP+-dependent morphine dehydrogenase that initiates morphine catabolism is encoded by a large plasmid of 165 kb. Treatment of P. putida M10 with ethidium bromide led to the isolation of a putative plasmid-free strain that was incapable of growth on morphine.


Read more at: Design of novel cationic ligands for the purification of trypsin‐like proteases by affinity chromatography

Design of novel cationic ligands for the purification of trypsin‐like proteases by affinity chromatography

A number on new cationic ligands have been designed and synthesized for the selective resolution an purification of the trypszin‐like proteases. A series of ligands based on 4‐[2′‐methyl‐4′‐(2″,4″‐dichloro‐1″,3″,5″‐triazin‐6‐ylamino) phenylazo]benzamidine were able to bind to trypsin and the trypsin‐like proteases, thrombin and urokinase, but bound pancreatic kallikrein only weakly.


Read more at: Acoustic Love plate sensors: a theoretical model for the optimization of the surface mass sensitivity

Acoustic Love plate sensors: a theoretical model for the optimization of the surface mass sensitivity


Read more at: Covalent Electropolymerization of Glucose Oxidase in Polypyrrole. Evaluation of Methods of Pyrrole Attachment to Glucose Oxidase on the Performance of Electropolymerized Glucose Sensors

Covalent Electropolymerization of Glucose Oxidase in Polypyrrole. Evaluation of Methods of Pyrrole Attachment to Glucose Oxidase on the Performance of Electropolymerized Glucose Sensors

Methods for covalently immobilizing glucose oxidase in polypyrrole are investigated. The enzyme was chemically modified with pyrrole using one of three different reactive side chains found in the protein. The reactions involve carbodiimide coupling to either lysyl or carboxyl residues on the enzyme and Schiff base reaction of the carbohydrate moiety. O ptimal coupling was achieved with the carbodiimide reaction, 15-20 mol of pyrrole/mol of enzyme compared with 6 mol of pyrrole/mol of enzyme for the Schiff base method.


Read more at: A Holographic Sensor for Proteases

A Holographic Sensor for Proteases

The use of holographic elements as biochemical sensors is suggested and exemplified with a device for monitoring protease activity. A quantitative optical response of a holographic element constructed in gelatin is demonstrated for a range of trypsin concentrations down to 25 nM with a response time within 20 min. These data demonstrate the principle for a general protease sensor which has particular relevance to the measurement of trypsin activity below normal physiological duodenal levels. The holographic devices respond with a change in wavelength (color) and/or a change in brightness.


Read more at: A hologram biosensor for proteases

A hologram biosensor for proteases

The concept of using a hologram as the interactive element in a one-shot biosensor is presented. The theoretical basis for a directly observed optical response to biological molecules is introduced. The most immediate application of such a device, restricted to the detection of proteases, is described in this paper. Using spectrographic measurements, a reflection hologram in gelatin in particular has been applied to the detection of 20 μg ml-1 trypsin and 23 μg ml-1 chymotrypsin, showing a greater sensitivity to trypsin.


Read more at: Noncontact excitation of high Q acoustic resonances in glass plates

Noncontact excitation of high Q acoustic resonances in glass plates

The applications of magnetic direct generation, a noncontact technique for generating acoustic waves, are limited by poor transduction efficiency. This letter describes an approach for substantially improving the transduction efficiency by application of a transduction format, enhanced direct magnetic generation, that utilizes continuous wave resonances and thin film generation. Glass plates coated with thin films of aluminium have been used as free-standing acoustic cavities to generate resonances with high transduction efficiencies and Q factors.