We have developed a rapid and sensitive thin film assay for in-process monitoring of target protein purification. This novel biosensor method provides rapid (5-min) visual evaluation of column purification fractions. The method can be used to monitor the efficiency of purification and potential loss of protein if the column binding capacity is exceeded. The eluted fractions containing the highe...

In this study Escherichia coli DE3 containing expression vector (pET21a) with cloned Persian sturgeon growth hormone (psGH) gene was grown in 10 mL LB broth on a 150 rpm shaker, at the temperature of 37 °C. At the late log phase (determined by OD standard curve) 100 &muL isopropyl &beta-D-1-thiogalactopyranoside (IPTG) was added for induction of GH synthesis. Samples were taken every 2 hours an...

background: streptokinase is a potent activator of plasminogen to plasmin, the enzyme that can solubilize the fibrin network in blood clots. streptokinase is currently used in clinical medicine as a thrombolytic agent. it is naturally secreted by β-hemolytic streptococci. methods: to reach an efficient method of purification, an immunoaffinity chromatography method was developed that could puri...

background: there are varieties of purification techniques for separation of human plasma proteins such as salting out, ion exchange chromatography, and ethanol fractionation. there are limitations for each method, for example in salting out method, the salt has to be removed in an additional step. ion exchange chromatography is difficult for scaling up, and plasma fractionation is a time consu...

in this study escherichia coli de3 containing expression vector (pet21a) with cloned persian sturgeon growth hormone (psgh) gene was grown in 10 ml lb broth on a 150 rpm shaker, at the temperature of 37 °c. at the late log phase (determined by od standard curve) 100 μl isopropyl β-d-1-thiogalactopyranoside (iptg) was added for induction of gh synthesis. samples were taken every 2 hours and afte...

The Protein Maker is an automated purification system developed by Emerald BioSystems for high-throughput parallel purification of proteins and antibodies. This instrument allows multiple load, wash and elution buffers to be used in parallel along independent lines for up to 24 individual samples. To demonstrate its utility, its use in the purification of five recombinant PB2 C-terminal domains...

background epidermal growth factor (egf) plays a fundamental role in the healing of wounds relating to skin damage, the cornea, and the gastrointestinal tract. objectives the aim of this study is the cloning, expression, and purification of recombinant human egf (rhegf), and an assessment of its activity. materials and methods in the present experimental study, a synthetic pet28a (+) -hegf cons...

Polymerase chain reaction (PCR) technique is widely used in many experimental conditions, and Taq DNA polymerase is critical in PCR process. In this article, the Taq DNA polymerase expression plasmid is reconstructed and the protein product is obtained by rapid purification, ("Rapid purification of high-activity Taq DNA polymerase" (Pluthero, 1993 [1]), "Single-step purification of a thermostab...

Most protein purification procedures include an affinity tag fused to either the N or C-terminal end of the protein of interest as well as a procedure for tag removal. Tag removal is not straightforward and especially tag removal from the C-terminal end is a challenge due to the characteristics of enzymes available for this purpose. In the present study, we demonstrate the utility of the divale...

Egg-laying hens were immunized with the Echis carinatus venom and the resulting antibodies were extracted from egg yolk by four different purification methods. The chicken egg yolk antibodies were purified by the water dilution method, polyethylene glycol (PEG) and ammonium sulphate precipitation method, chloroform extraction and the Lithium sulphate precipitation method. These methods were com...