Research for the production of recombinant human epidermal growth factor using Samia Cynthia Ricini pupae bioreactor / 生物医学工程学杂志

The protein production system using a baculovirus Antheraea pernyi nucleopolyhedrovirus (AnpeNPV) as a gene expression vector and its host insect as a natural bioreactor was successful established and its excellent performance in the protein production has been demonstrated. In this paper, the system is used to produce recombinant human epidermal growth factor (rhEGF), which have been widely used in medical and cosmetic treatment. A recombinant AnpehEGF virus has been constructed by replacing the viral polyhedrin gene with the rhEGF gene, and then injected it to Samia cynthia ricini pupae. Amplification and expression of rhEGF gene in the pupae was clearly detected by PCR, Western blot and ELISA analyses. These analyses have also revealed that rhEGF in the pupae was significantly increased at 6 days post-infection, and reached maximum level at the 12th day. The concentrations of rhEGF were 19.77, 24.90, 618.59 and 1 952.46 ng/g pupae at 3, 6, 9 and 12 days post-infection, respectively. However, the rhEGF concentration reduced at later stage (days 15). The rhEGF in the pupae could be purified using ammonium sulfate precipitation and Ni-NTA agrose affinity chromatography. Results demonstrate that Samia cynthia ricini pupae can be used as a bioreactor to produce rhEGF and, if successfully improved, will be a novel method of rhEGF production with lower cost and more efficient.

Comparison of serum protein profiles between people with successful aged 80 years and over in Bama versus Nanning city / 中华老年医学杂志

Objective To compare the serum protein profiles between people with successful aged 80 years and over in Bama versus Nanning,to identify the possible causes of the long life expectancy in Bama.Methods We evaluated the serum protein profiles using surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF-MS) with 4 different chromatographic surfaces,including the weak cation exchange (CM10),immobilized metal affinity capture (IMAC) 30-Cu,strong anion exchange (Q10) and hydrophobic surface (H50) protein chip arrays in people with successful aged 80 years and over living in Bama versus in Nanning city.SELDI-TOF-MS had a molecular mass detection range of 2000-50,000 D,with a first signal-to noise (S/N) ratio of 5,frequency of peak cluster of over 15 ％,and second S/N ratio of 2 and peak fluctuation of 0.3％ (m/z).Results The total numbers of serum protein (peptide) peaks for people with successful aging in Bama and Nanning were 71 on CM10,46 on IMAC30-Cu,81 on Q10,and 52 on H50.Among subjects from Bama as compared with Nanning (P＜0.05; mean ＞SD for each protein peak),there were 8 differently expressed protein peaks on CM10,of which 2 were upregulated and 6 downregulated; 7 on IMAC-Cu,of which 2 were upregulated and 6 downregulated; 5 on Q10,all of which were upregulated; and 6 on H50,of which 3 were upregulated and 3 downregulated.Conclusions There are marked differences in the serum protein profiles between people with successful aging living in Bama versus in Nanning.Most of the proteins are downregulated among people aged over 80 years in Bama.