AI Article Synopsis
Article Abstract
Pichia pastoris is a methylotropic yeast that has gained great importance as an organism for protein expression in recent years. Here, we report the expression of recombinant human erythropoietin (rhEPO) in glycoengineered P. pastoris. We show that glycosylation fidelity is maintained in fermentation volumes spanning six orders of magnitude and that the protein can be purified to high homogeneity. In order to increase the half-life of rhEPO, the purified protein was coupled to polyethylene glycol (PEG) and then compared to the currently marketed erythropoiesis stimulating agent, Aranesp(®) (darbepoetin). In in vitro cell proliferation assays the PEGylated protein was slightly, and the non-PEGylated protein was significantly more active than comparator. Pharmacodynamics as well as pharmacokinetic activity of PEGylated rhEPO in animals was comparable to that of Aranesp(®). Taken together, our results show that glycoengineered P. pastoris is a suitable production host for rhEPO, yielding an active biologic that is comparable to those produced in current mammalian host systems.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.jbiotec.2011.11.002 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Scorpion insect neurotoxin LqhIT2 is a promising oral biopesticide: high-level preparation in Pichia pastoris and bioactivity assays.
Pest Manag Sci
December 2024
Postdoctoral Mobile Station of Biology, Genetic Engineering Research Center, College of Life Sciences, Chongqing University, Chongqing, China.
Background: Discovering insecticidal proteins with high activity and strict insect specificity and applying them to the biological control of insect pests is of great significance. Oral LqhIT2 has insecticidal activity, which most other insecticidal neurotoxin proteins do not have, but the large-scale preparation of the toxin is difficult and one of the obstacles to determining its anti-insect potential for biological control.
Results: In this study, the expression level of recombinant LqhIT2 (rLqhIT2) in Pichia pastoris was as high as 1.
Modulating the pH-activity profile of the glucose isomerase from Thermotoga marimita by introducing positively and negatively charged residues.
Biophys Chem
December 2024
School of Food Science and Engineering, Ocean University of China, Qingdao, Shandong 266500, China.
Glucose isomerase is generally used in the industrial production of high-fructose corn syrup, and a heat- and acid-resistant glucose isomerase is preferred. However, most glucose isomerases exhibit low activity or inactivation at low pH. In this study, we demonstrated that two combination mutants formed by introducing positive and negative charges near the active site and on the surface of the enzyme demonstrated a successful reduction in the optimal pH and increase in the specific activity of glucose isomerase from Thermotoga maritima (TMGI).
View Article and Find Full Text PDFProduction of virus-like particles of FMDV by 3C protease cleaving precursor polyprotein P1 in vitro.
Appl Microbiol Biotechnol
December 2024
State Key Laboratory for Animal Disease Control and Prevention, College of Veterinary Medicine, Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730000, China.
Nonstructural protein 3C, a master protease of Picornaviridae, plays a critical role in viral replication by directly cleaving the viral precursor polyprotein to form the viral capsid protein and antagonizing the host antiviral response. Additionally, 3C protease, as a tool enzyme, is involved in regulating polyprotein expression. Here, the 3C mutant gene (3Cm), fused with a small ubiquitin-like modifier (SUMO) tag at the N-terminal and featuring a mutation at position 127, was inserted into the cold-shock plasmid pCold of Escherichia coli for expression.
View Article and Find Full Text PDFProtein adsorption to poly(ethylenimine)-modified Sepharose FF: IX. Further studies on counterion effects and behavior in therapeutic protein separation.
J Chromatogr A
December 2024
Department of Biochemical Engineering, School of Chemical Engineering and Technology and Key Laboratory of Systems Bioengineering and Frontiers Science Center for Synthetic Biology (Ministry of Education), Tianjin University, Tianjin 300350, PR China. Electronic address:
Our previous studies on protein adsorption onto anion-exchangers of poly(ethylenimine) (PEI)-grafted Sepharose FF (PEI-Sepharose) proved their significantly improved performance over the commercial nongrafting anion-exchangers such as Q Sepharose FF, and it was found the protein adsorption behavior on PEI-Sepharose was more sensitive to counterions (Cl, SCN, HPO and SO). However, the complicated role of counterions has not been well interpreted due to their distinct chemical and physical characteristics. Thus, we have further studied the counterion effects by adding two halide ions (F and Br) to explore the effects of the three halide ions on bovine serum albumin adsorption and the results were compared with previous data.
View Article and Find Full Text PDFMolecular mechanism of GSH metabolism and autophagy in NAC-promoted recombinant human serum albumin and follicle stimulating hormone beta fusion protein secretion in Pichia pastoris.
J Biotechnol
December 2024
Jiangsu Key Laboratory of Sericultural and Animal Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, China; Key Laboratory of Silkworm and Mulberry Genetic Improvement, Ministry of Agriculture and Rural Affairs, Sericultural Scientific Research Center, Chinese Academy of Agricultural Sciences, Zhenjiang 212100, China. Electronic address:
The Pichia pastoris expression system is a favorable platform for production of pharmaceutical proteins. Treatment of strains with N-acetyl-L-cysteine (NAC) has been shown to enhance the yield of recombinant proteins, thereby contributing to a reduction in production costs. However, the specific mechanism of action of NAC remains unclear.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!