EPO a Human, Sf9
Description
EPO-a is expressed with an 8 amino acid His tag at C-Terminus and purified by proprietary chromatographic techniques.
Product Specs
Product Science Overview
Erythropoietin (EPO) is a glycoprotein hormone that plays a crucial role in the regulation of erythropoiesis, the process by which red blood cells (erythrocytes) are produced. It is primarily produced in the kidneys in response to hypoxia (low oxygen levels) and acts by binding to the erythropoietin receptor (EPO-R) on the surface of erythroid progenitor cells in the bone marrow, stimulating their proliferation and differentiation into mature red blood cells .
Recombinant human erythropoietin (rHuEPO), such as Erythropoietin-alpha, is a biotechnologically engineered form of the natural hormone. It is produced using recombinant DNA technology, which involves inserting the human erythropoietin gene into host cells to produce the protein. One of the host systems used for this purpose is the Sf9 insect cell line, derived from the fall armyworm (Spodoptera frugiperda). This system is advantageous due to its ability to perform post-translational modifications similar to those in mammalian cells, which are essential for the biological activity of the protein .
Erythropoietin-alpha is a 30.4 kDa glycoprotein composed of 165 amino acids arranged in four antiparallel alpha helices. It contains two beta sheet bonds and two intrachain disulfide bridges at positions Cys7–Cys161 and Cys29–Cys33. The carbohydrate portion of the molecule accounts for approximately 40% of its molecular weight and includes N-linked glycosylation at asparagine residues 24, 38, and 83, as well as O-linked glycosylation at serine residue 126 .
The glycosylation of EPO is critical for its stability, solubility, and biological activity. It protects the protein from proteolytic degradation and modulates its interaction with the EPO receptor. The degree of sialylation (attachment of sialic acid residues) influences the protein’s half-life in circulation, with highly sialylated forms having a longer half-life .
Recombinant erythropoietin-alpha is widely used in clinical settings to treat various forms of anemia, particularly those associated with chronic kidney disease, cancer chemotherapy, and certain HIV treatments. By stimulating erythropoiesis, it helps to increase red blood cell counts and improve oxygen delivery to tissues, thereby alleviating symptoms of anemia such as fatigue and weakness .
In addition to its erythropoietic effects, erythropoietin-alpha has been shown to have neuroprotective and neurotrophic properties. It can confer protection to neurons in various models of neurological diseases, including stroke and neurodegenerative disorders. This has led to ongoing research into its potential therapeutic applications beyond hematology .
The production of recombinant erythropoietin-alpha in Sf9 cells involves several steps:
- Gene Cloning: The human erythropoietin gene is cloned into a suitable expression vector.
- Transfection: The vector is introduced into Sf9 cells, which are then cultured to express the recombinant protein.
- Protein Expression: The cells produce and secrete erythropoietin-alpha into the culture medium.
- Purification: The protein is purified from the culture medium using techniques such as affinity chromatography, which exploits the specific binding properties of the protein to isolate it from other cellular components .
Despite its success, the production of recombinant erythropoietin-alpha faces several challenges, including the need for efficient purification methods and the potential for immunogenicity (immune response against the protein). Advances in biotechnology continue to address these issues, with ongoing research focused on optimizing production processes and exploring new therapeutic applications for erythropoietin and its derivatives .
