Epoetin beta

Manufactured by Roche

Epoetin beta is a recombinant human erythropoietin (rHuEPO) used as a pharmaceutical agent. It is a glycoprotein that stimulates the production of red blood cells in the bone marrow.

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Lab products found in correlation

Dithiothreitol (dtt), by Merck Group (3 mentions) Sodium phosphate, by Merck Group (2 mentions) Monobasic and dibasic, by Merck Group (2 mentions) Asp n, by Roche (2 mentions) Pngase f, by Roche (2 mentions) Glu c, by Roche (2 mentions) Trypsin, by Roche (1 mentions) Guanidine hcl, by Merck Group (1 mentions) Sequencing grade trypsin, by Promega (1 mentions) Iodoacetamide, by Merck Group (1 mentions) Urea, by Merck Group (1 mentions) Sialidase, by Roche (1 mentions) Formic acid, by Merck Group (1 mentions) Acetonitrile acn, by Biosolve (1 mentions)

4 protocols using epoetin beta

Erythropoietin BRP Characterization

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EPO BRP (EDQM, Ph. Eur.) contains a freeze-dried preparation of erythropoietin in vials with a declared content of 250μg of erythropoietin per vial. This erythropoietin BRP is known as a 50:50 mixture of epoetin alfa (The R.W. Johnson pharmaceutical Research Institute) and epoetin beta (Boehringer-Mannheim, recently Roche).
Trypsin, Asp-N, Glu-C, and PNGaseF were purchased from Roche. Sodium phosphate, monobasic and dibasic, DTT, ad guanidine-HCl were purchased from Sigma. Acetonitrile (ACN), trifluoacetic acid (TFA) and methanol were purchased from B&J. Acetic acid was purchased from J.T. Baker.

Byeon J., Lim Y.R., Kim H.H, & Suh J.K. (2015). Structural Identification of a Non-Glycosylated Variant at Ser126 for O-Glycosylation Site from EPO BRP, Human Recombinant Erythropoietin by LC/MS Analysis. Molecules and Cells, 38(6), 496-505.

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Characterization of Erythropoietin BRP

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EPO BRP (EDQM, Ph. Eur.) contains a freeze-dried preparation of erythropoietin in vials with a declared content of 250 μg of erythropoietin per vial. This erythropoietin BRP is known as a 50:50 mixture of epoetin alfa (The R.W. Johnson pharmaceutical Research Institute) and epoetin beta (Boehringer-Mannheim, recently Roche).
Endoproteinase trypsin was purchased from Roche. Sodium phosphate, monobasic and dibasic, DTT, and guanidine-HCl were purchased from Sigma. Acetonitrile (ACN), TFA and methanol were purchased from B&J. Acetic acid was purchased from J.T. Baker.

Song K.E., Byeon J., Moon D.B., Kim H.H., Choi Y.J, & Suh J.K. (2014). Structural Identification of Modified Amino Acids on the Interface between EPO and Its Receptor from EPO BRP, Human Recombinant Erythropoietin by LC/MS Analysis. Molecules and Cells, 37(11), 819-826.

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Erythropoietin Impacts Tumor Growth in Nude Mice

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Mouse experiments were performed in accordance with the Swiss animal law and with the approval of the ethical committee of the respective local authorities (Kanton Zurich). Hsd : Athymic Nude-Foxn1^nu (8-9 weeks old) male mice (Envigo, Netherlands) were kept in a pathogen-free mouse barrier facility (22 ± 5°C in a 12 h light/dark cycle; standard rodent chow (Kliba Nafag, #3436) and water ad libitum). We injected 3x10⁶ cells in a 1:6 Matrigel-PBS solution into the rear right flank. Five different cell lines, namely wild-type, shSCR1, shSCR2, shEPOR1, and shEPOR2 cells, were injected into groups of 16 nude mice. Seven days after tumor cell injection, mice were cage-wise split into two groups of eight animals. Subsequently, eight mice were intraperitoneally injected with 300 U/kg EPO (Epoetin-beta; Recormon^®, Roche) (6 (link), 25 (link)) and the other eight mice were injected with saline throughout the entire experiment. Additional confounders were not identified and controlled for. Tumor size was calculated V=1/2*Length*(Width)² after measuring the length (largest tumor diameter) and width (perpendicular tumor diameter) with a caliper (26 (link)). Mice were euthanized with CO₂ and blood for hemoglobin measurements and hematocrit (ABL 800 Flex, Radiometer) was retrieved from the right heart ventricle.

Aboouf M.A., Guscetti F., von Büren N., Armbruster J., Ademi H., Ruetten M., Meléndez-Rodríguez F., Rülicke T., Seymer A., Jacobs R.A., Schneider Gasser E.M., Aragones J., Neumann D., Gassmann M, & Thiersch M. (2022). Erythropoietin receptor regulates tumor mitochondrial biogenesis through iNOS and pAKT. Frontiers in Oncology, 12, 976961.

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Recombinant erythropoietin characterization

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The recombinant rhEPO BRP (presently batch 4.0) was acquired from EDQM (European Directorate for the Quality of Medicines). It is a 50:50 blending of epoetin alpha and Epoetin beta53 , supplied at 13,000 IU per vial. Epoetin beta was acquired from Roche and supplied at 30,000 IU per syringe. It is a recombinant form of erythropoietin marketed under the trade name NeoRecormon, produced by recombinant DNA technology using Chinese hamster ovary cells as expression system. Epoetin zeta (Retacrit, Hospira) is a biosimilar to the reference medicinal product Eprex (Epoetin alpha), produced in Chinese hamster ovary cells. The Uniprot code for rhEPO is P01588. Human blood plasma properdin (Uniprot code: P27918) was obtained from Complement Technology, Inc. (TX, USA). Urea, dithiothreitol (DTT) and iodoacetamide were purchased from Sigma-Aldrich (Steinheim, Germany). Formic acid (FA) was purchased from Merck (Darmstadt, Germany). Acetonitrile (ACN) was purchased from Biosolve (Valkenswaard, The Netherlands). Sequencing grade trypsin was obtained from Promega (Madison, WI, USA). Glu-C, Asp-N, PNGase F and Sialidase were obtained from Roche (IN, USA).

Yang Y., Liu F., Franc V., Halim L.A., Schellekens H, & Heck A.J. (2016). Hybrid mass spectrometry approaches in glycoprotein analysis and their usage in scoring biosimilarity. Nature Communications, 7, 13397.

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