Protein purification is often a bottleneck during protein generation for large molecule drug discovery. Therapeutic antibody campaigns typically require the purification of hundreds of monoclonal antibodies (mAbs) during the hybridoma process and lead optimization. With the increase in high-throughput cloning, faster DNA sequencing, and the use of parallel protein expression systems, a need for...

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: 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...

Homeobox genes encode transcription factors which play important roles in the developmental processes of many multicellular organisms. TGIFLX/Y (TGIFLX and TGIFLY) are members of the homeobox superfamily of genes. Their expressions are specifically detected in the human adult testis but their functions are remained to be investigated. In this investigation we cloned full length of TGIFLY cDNA a...

background:the dynamic binding capacity (dbc) of a chromatography matrix in protein purification is the amount of the total protein absorbed into the matrix, before occurrence of a significant break in the breakthrough curve. optimization of the process criteria for maximum dbc avoids extra process scale-up and reduces the processing time, costs and protein loss. taguchi method is a simple usef...

In the process of protein purification, the amount of proteins isolated with the help of commercial protein purification processes remains uncertain and vague. The present paper proposed a set of fuzzy rule system based on Fuzzy Expert System (FES) which predicts the amount of purified proteins based upon the flow rate of the protein in column, pH and binding capacity of resin to desired protei...

This paper describes methods to produce an isotopically labeled 23 kDa viral membrane protein with purified yield of 20 mg/L of Esch­e­rich­ia coli shake flask culture. This yield is suf­fi­cient for NMR structural studies and the protein production methods are simple, straightforward, and rapid and likely applicable to other recombinant membrane proteins expressed in E. coli. The target FHA2 p...

Article history: Received 5 December 2007 and in revised form 12 March 2008 Available online 25 March 2008

BACKGROUND Recombinant protein expression and purification remains a fundamental issue for biotechnology. Recently we found that two short self-assembling amphipathic peptides 18A (EWLKAFYEKVLEKLKELF) and ELK16 (LELELKLKLELELKLK) can induce the formation of active protein aggregates in Escherichia coli (E. coli), in which the target proteins retain high enzymatic activities. Here we further exp...