• Biomolecules & Therapeutics
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• v.2 no.1
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• pp.77-81
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• 1994

The acute toxicity of recombinant human epidermal growth factor, DWP-401 was evaluated in ICR mice of both sexes. Six groups of mice were administered orally or subcutaneously with 0, 0.125, 0.25, 0.5, 1 and 2 mg/kg of DWP-401. Abnormal clinical signs related to the compound were not observed, and no deaths occurred. Gross findings of necropsy revealed no evidence of specific toxicity related to DWP-401. LD$_{50}$ values for both male and female mice were evaluated to be over 2 mg/kg, which is approximately 2, 000 fold of presumed clinical dose.e.

• Journal of Food Hygiene and Safety
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• v.9 no.1
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• pp.31-36
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• 1994

The acute toxicity of recombinant human epidermal growth factor, DWP-401 was evaluated in SD rats. Male and female rats aging 6 weeks were administered orally or subcutaneously with 0, 0.125, 0.25, 0.5, 1 and 2 mg/kg of DWP-401. No deaths and no toxic symptoms related to the DWP-401 were observed. The body weights of treated animals were not significantly different from the controls. The results of necropsy revealed no abnormal gross findings of the organs in treated animals. LD50 values of DWP-401 for male and female rats were estimated to be over 2 mg/kg, which is approximately 2, 000 times of expected clinical dose.

• Biomolecules & Therapeutics
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• v.4 no.2
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• pp.138-147
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• 1996

DWP401, a recombinant human epidermal growth factor, was subcutaneously administered to ICR mice at the dose levels of 0, 0.04, 0.2 and 1.0 mg/kg/day (15rats/sex/group) in order to evaluate the subchronic toxicity. General observations, examinations for food and water consumption, ophthalmoscopy and urinalysis were carried out during the study. For the complete gross and microscopic examinations, 10 mice/ sex/group were sacrificed at the ends of the dosing period, and the remaining animals were sacrificed with a 5 week recovery period. Examinations for hematology and blood biochemistry were also carried out at the time of recovery period. Based on the results, it was thought that the target tissue or organs were mesothelial cell, injection site, spleen, adrenal gland, ovary and transitional epithelial cell of urinary tract, and no observed toxic level of DWP401 was 0.04 mg/kg while definite toxic dose level might be 0.2 mg/kg.

• Archives of Pharmacal Research
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• v.26 no.8
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• pp.649-652
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• 2003

Polymeric nanoparticles composed of polystyrene (PS) as core and poly(methacrylic acid) (PMA) as corona were prepared by the dispersion copolymerization. The potential of the nanoparticles as carriers for recombinant human epidermal growth factor (EGF) was investigated. The nanoparticles showed monodispersity and good water-dispersibility. The loading content of EGF to the nanoparticles was very high due to electrostatic interaction between EGF and nanoparticles. EGF was released as a pseudo-zero order pattern after initial burst effect. The nanoparticles were sufficient for A431 cells proliferation.

• Biotechnology and Bioprocess Engineering:BBE
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• v.2 no.2
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• pp.86-89
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• 1997

Epidermal growth factor(EGF) known as a urgastrone is a powerful mitogen with a wide variety of possibilities for medical usages. A mature EGF coding region was isolated from human prepro-EGF sequence by a conventional PCR and cloned into pQE vector in which the gene product was supposed to be expressed with 6$\times$His tag for the subsequent purification. The recombinant mature EGF was expressed in M15[Rep4], an Escherichia coli host strain, in amount of 30-40% of total proteins pressent in E. coli extract by the addition of isopropylthio-$\beta$-galactopyranoside (IPTG). The recombinant EGF purified using a Ni2+-NTA affinity colume chromatography was active in its ability to induce phosphorylation on tyrosine residues of several substrate proteins when murine NH3T3 and human MRC-5 fibroblast cells were stimulated with it. This work may provide the basic technology and information for the production of recombinant EGF.

• Archives of Pharmacal Research
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• v.24 no.4
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• pp.355-359
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• 2001

A novel HPLC method with electrochemical detection has been developed for the determination of recombinant human epidermal growth factor (rhEGF) in pharmaceutical products. rhEGF was separated from other components in formulation on a reversed-phase C18 column with 24% acetonitrile in 0.1 M phosphate buffer (pH 4.75). The optimum electrochemical oxidation of EGF was obtained at 0.85 V vs. Ag/AgCl in a glassy carbon working electrode due to electroactive tyrosine, tryptophan, methionine, and arginine residues. The quantitation range was from 1.0 to 200 ng of rhEGF with the linear correlation coefficient greater than 0.999. The method was successfully applied for the quantitation of rhEGF in a pharmaceutical preparation.

• Archives of Pharmacal Research
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• v.24 no.6
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• pp.601-606
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• 2001

A simple assay method of recombinant human epidermal growth factor (rhEGF) in a pharmaceutical preparation was studied and validated by capillary electrophoresis (CE) using micellar electrokinetic chromatography (MEKC) techniques. Factors affecting the migration behavior and separation performances of the peptide; type of buffers pH, butler concentration, and concentration of sodium dodecyl sulfates (SDS) were investigated to optimize the analytical performance. CE was performed using running buffers 50.0 mM borate (pH 8.5) containing 12.5 mM SDS at 20 $mutextrm{V}$ of the applied voltage. Calibration curves for the rhEGF showed good linearity (r>0.999) over the wide dynamic range from 1.25 to $100{\mu\textrm{g}}/ml$. Sample analysis was performed by using standard addition method to eliminate the matrix effects of dosage vehicle. This method is assumed to be useful for quality control (QC) of various forms of pharmaceutical products of the peptide.

• Biomedical Science Letters
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• v.6 no.2
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• pp.83-91
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• 2000

Subconfluent neonatal human epidermal keratinocytes were treated with a concentration 200 ng/$m\ell$ of human recombinant epidermal growth factor (hrEGF), human recombinant insulin-like growth factor-1 (hrIGF-1), and with a combination of hrEGF and hrIGF-1. Cytoplasmic and membrane-associated proteins were extracted and assayed. Proteins were separated by SDS-PAGE, and subjected to the western blot analysis. In the cytoplasmic fraction, the PKC concentration of keratinocyte treated with hrIGF-1 was higher than the control group, but the concentration of control group was the highest than the others in the membrane fraction. In the cytoplasmic fraction, EGF stimulated PKC-$\beta$II, -$\delta$, -$\theta$, and also stimulated PKC-$\alpha$, -$\beta$I, -$\delta$, -$\Im$ and -$\theta$ in the membrane fraction. IGF-1 stimulated PKC-$\beta$I, -$\Im$ and -$\theta$ in the cytoplasmic, PKC-$\alpha$, -$\beta$I, -$\delta$, -$\Im$, - $\varepsilon$ and -$\theta$ in the membrane. In the cells treated with a combination of EGF and IGF-1, PKC-$\alpha$, -$\beta$I, -$\Im$ and -$\theta$ in the cytoplasmic fraction, PKC-$\alpha$, -$\delta$, -$\Im$, -$\varepsilon$ and -$\theta$ in the membrane fraction were stimulated.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.299.1-299.1
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• 2003

Multivesicular DepoFoam technology is best suited for the encapsulation and sustained release of water-soluble drugs. The purpose of the present study was to prepare multivesicular DepoFoam particles and investigated possibility of oral delivery of a peptide, human epidermal growth factor (rhEGF). The multivesicular DepoFoam particles containing rhEGF was prepared by a two step water-in-oil-in-water double emulsification process. (omitted)

• Microbiology and Biotechnology Letters
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• v.24 no.4
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• pp.478-484
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• 1996

Recombinant human epidermal growth factor (rhEGF) was produced by E. coli BL21 harboring a plasmid pYHB101. The maximum production was 68.7 mg/l when the E. coli strain was cultured at 25$\circ$C for 48 hours in the modified MBL medium containing 10 g/l glucose with 1 mM IPTG induction at 2 hours after inoculation. The rhEGF was purified upto 267 folds by Amberlite XAD- 7 chromatography, ultrafiltration, and DEAE Sepharose fast flow ion exchange chromatography with an overall yield of 66.6%. The purified rhEGF was further separated into two fractions by HPLC. The N-terminal amino acid sequence of the second fraction was Asn-Ser-Asp-Ser-Glu-Cys-Pro-Leu-Ser-His. The effect of rhEGF on the DNA synthesis was examined using in vitro biological assay based on the incorporation of 5'-bromo-2'- deoxy-uridine (BrdU). The purified rhEGF shows no difference with natural human epidermal growth factor (nhEGF) in N-terminal amino acids residues and biological activity. From the results, we concluded that rhEGF produced from E. coli harboring the plasmid pYHB101 was apparently the same as nhEGF.