High level expression of human leukemia inhibitory factor (LIF) from a synthetic gene in Escherichia coli and the physical and biological characterization of the protein

A simple method for production and purification of soluble and biologically active recombinant human leukemia inhibitory factor (hLIF) fusion protein in Escherichia coli

“…The hLIF fusion proteins purities were greater than 95% (Table 1). The yields of His 6 -hLIF and Trx-His 6 -hLIF fusion proteins in this work were higher than previously reported (Gearing et al, 1989;Samal et al, 1995;Tomala et al, 2010).…”

“…3B), corresponding to a specific activity of 1.7 × 10 7 , 1.3 × 10 7 and 1.5 × 10 7 units per mg, respectively. These specific activities are similar to other recombinant hLIF preparations (Gearing et al, 1989;Samal et al, 1995). This study has shown that the His 6 -hLIF and Trx-His 6 -hLIF fusion proteins were effective in inducing TF-1 cells proliferation, indicating that the presence of the His 6 or TrxHis 6 at the N-termini of the hLIF fusion proteins do not interfere with their biological functions.…”

“…The final yield of hLIF fusion proteins exceeded that obtained from previous studies (Gearing et al, 1989;Samal et al, 1995;Tomala et al, 2010). The biological activities of these purified proteins are comparable to commercial hLIF (Fig.…”

“…Many studies have successfully used Escherichia coli to produce several human growth factors and cytokines, such as interleukin-1 (Furutani et al, 1985), interleukin-4 (van Kimmenade et al, 1988), epidermal growth factor (Shimizu et al, 1991;Abdull Razis et al, 2008), fibroblast growth factor (Squires et al, 1988;Gasparian et al, 2009) and LIF (Gearing et al, 1989;Samal et al, 1995;Tomala et al, 2010) for therapeutic and clinical purposes. Although, recombinant human LIF (hLIF) is available and has been successfully produced in E. coli expression systems, they still require many steps of downstream processing.…”

Preparation of bioactive soluble human leukemia inhibitory factor from recombinant Escherichia coli using thioredoxin as fusion partner

“…Gearing et al [12] expressed murine and human LIF in a glutathione-Stransferase based fusion construct while Samal et al [13] reported the expression of mature human LIF as insoluble inclusion bodies with subsequent refolding procedures. In this work we describe the development of a novel production and purification process for recombinant human LIF (hLIF).…”