Expression of Recombinant Human Epidermal Growth Factor in Escherichia coli and Characterization of its Biological Activity

Abstract

Recombinant human epidermal growth factor (EGF) was successfully expressed as a fusion protein in Escherichia coli system. This system was used OmpA signal sequence to produce soluble protein into the periplasm of E. coli. Human EGF (hEGF) synthesized in bacterial cell was found to be similar in size with the original protein and molecular weight approximately at 6.8 kDa. Cell proliferation assay was conducted to characterize the biological activity of hEGF on human dermal fibroblasts. The synthesized hEGF was found to be functional as compared with authentic hEGF in stimulating cell proliferation and promoting growth of cell. In comparison of biological activity between synthesized and commercial hEGF on cell proliferation, the results showed there was no significant different. This finding indicates the synthesized hEGF in E. coli system is fully bioactive in vitro.

Genetically Engineered Epidermal Growth Factor Conjugate Crosses Cell Membrane

“…EGF exists in many tissues and organs [18] as well as various body fluids [17]. EGF can stimulate cellular proliferation [11], leading to the generation of types of cells [3], and exhibit overlapping functions with other trophic factors and cytokines [1]. EGF is also a potent differentiation factor for neural progenitors and stem cells [2,11].…”

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

“…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.…”

Cloning, Expression, and Cost Effective Purification of Authentic Human Epidermal Growth Factor With High Activity

“…A pilot experiment was performed using a cell concentration of 1 × 10 4 cells/well in a 96-well microculture plate. The expressed rhEGF and commercial EGF were added to the wells (100 μL) in 4 different concentrations of EGF (1, 10, 100, and 500 ng/mL) ( 12 , 17 ). Control experiments were carried out using the complete growth culture medium without rhEGF.…”