Recombinant human granulocyte-macrophage colony-stimulating factor: effect of glycosylation on pharmacokinetic parameters

Marini, Guillermo; Forno, Guillermina; Kratje, Ricardo; Etcheverrigaray, Marina

doi:10.2225/vol10-issue2-fulltext-5

Cited by 8 publications

(8 citation statements)

References 10 publications

(12 reference statements)

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“…It is a general fact that quantity and quality of the recombinant protein as well as the cost of production are important considerations in producing pharmaceutical therapies. The recombinant proteins which are produced by different host cells vary in protein characteristic such as effective translation due to codon specificity, post-translational modifications, as well as the size of the resulting protein which is dependent on the degree of glycosylation [ 8 ]. For instance, certain recombinant proteins expressed in E. coli required extra processing steps for denaturation and refolding, in order to obtain biologically active forms [ 18 ].…”

Section: Discussionmentioning

confidence: 99%

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A Comparative Study on the Expression, Purification and Functional Characterization of Human Adiponectin in Pichia pastoris and Escherichia coli

Rothan

Teh

Haron

et al. 2012

IJMS

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Adiponectin is one of the most bioactive substances secreted by adipose tissue and is involved in the protection against metabolic syndrome, artherosclerosis and type II diabetes. Research into the use of adiponectin as a promising drug for metabolic syndromes requires production of this hormone in high quantities considering its molecular isoforms. The objective of this study is to produce recombinant human adiponectin by Pichia pastoris (P-ADP) as a cheap and convenient eukaryotic expression system for potential application in pharmaceutical therapy. For comparison, adiponectin was also expressed using the Escherichia coli (E-ADP) expression system. Adiponectin was constructed by overlap-extension PCR, and cloned in standard cloning vector and hosts. Recombinant expression vectors were cloned in the P. pastoris and E. coli host strains, respectively. SDS-PAGE and western blotting were used to detect and analyse expressed recombinant protein in both systems. Adiponectin was purified by affinity chromatography and quantified using the Bradford Assay. The results of this study indicated that P-ADP quantity (0.111 mg/mL) was higher than that of E-ADP (0.04 mg/mL) and both were produced in soluble form. However, P-ADP was able to form high molecular weights of adiponectin molecules, whilst E-ADP was not able to form isoforms higher than trimer. In addition, P-ADP was more active in lowering blood glucose compared with E-ADP. The two types of proteins were equally efficient and significantly decreased blood triglyceride and increased high density lipoprotein. We conclude that P. pastoris is able to produce high quantity of bioactive adiponectin for potential use in treatment of metabolic syndromes.

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Section: Discussionmentioning

confidence: 99%

“…It is generally accepted that the choice of heterologous protein expression system has profound influence on characteristics of the recombinant protein [ 8 ]. Compared to prokaryotic expression systems like Escherichia coli , the advantages of eukaryotic expression system such as the methylotrophic yeast Pichia pastoris are many.…”

Section: Introductionmentioning

confidence: 99%

A Comparative Study on the Expression, Purification and Functional Characterization of Human Adiponectin in Pichia pastoris and Escherichia coli

Rothan

Teh

Haron

et al. 2012

IJMS

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“…delivery increased the half-life of recombinant glycosylated GMCSF to 25 h when the half-life was only 3 min when administered i.v. (45). A disadvantage of changing the route of administration is that maintaining the effective concentration of the drug at the tumor may not be possible.…”

Section: Pharmacokinetics and Pharmacodynamicsmentioning

confidence: 99%

Turning the gene tap off; implications of regulating gene expression for cancer therapeutics

Curtin

Candolfi

Xiong

et al. 2008

Molecular Cancer Therapeutics

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“…Glycosylation can influence pharmacokinetics and protein function, so changes in the glycosylation profile can have a significant impact on the pharmacodynamic characteristics and consequently therapeutic efficacy (Sinclair and Elliot, 2004;Marini et al, 2005).…”

Section: Carbohydrate Determinationmentioning

confidence: 99%

Animal cell technology: from biopharmaceuticals to gene therapy

2008

Choice Reviews Online

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Recombinant human granulocyte-macrophage colony-stimulating factor: effect of glycosylation on pharmacokinetic parameters

Cited by 8 publications

References 10 publications

A Comparative Study on the Expression, Purification and Functional Characterization of Human Adiponectin in Pichia pastoris and Escherichia coli

A Comparative Study on the Expression, Purification and Functional Characterization of Human Adiponectin in Pichia pastoris and Escherichia coli

Turning the gene tap off; implications of regulating gene expression for cancer therapeutics

Animal cell technology: from biopharmaceuticals to gene therapy

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