<abstract xmlns="http://www.w3.org/1999/xhtml"><p>The aim of this study was to optimize the formulation of erythropoietin (EPO) using amino acids instead of human serum albumin (HSA) and to evaluate its <italic>in vivo</italic> stability in order to avoid the risk of viral contamination and antigenicity. Different EPO formulations were developed in such a way as to allow studying the effects of amino acids and surfactants on the EPO stability profile. The main techniques applied for EPO analysis were ELISA, Bradford method, and SDS gel electrophoresis. The <italic>in vivo</italic> stability was evaluated in a Balb-c mouse animal model. The results showed that the presence of surfactant was very useful in preventing the initial adsorption of EPO on the walls of vials and in minimizing protein aggregation. Amino acid combinations, glycine with glutamic acid, provided maximum stability. Formulation F4 (containing glycine, glutamic acid and Tween 20) showed minimum aggregation and degradation and <italic>in vivo</italic> activity equivalent to commercially available HSA-stabilized EPO (Eprex<sup>®</sup>).</p></abstract>

The aim of this study was to optimize the formulation of erythropoietin (EPO) using amino acids instead of human serum albumin (HSA) and to evaluate its in vivo stability in order to avoid the risk of viral contamination and antigenicity. Different EPO formulations were developed in such a way as to allow studying the effects of amino acids and surfactants on the EPO stability profile. The main techniques applied for EPO analysis were ELISA, Bradford method, and SDS gel electrophoresis. The in vivo stability was evaluated in a Balb-c mouse animal model. The results showed that the presence of surfactant was very useful in preventing the initial adsorption of EPO on the walls of vials and in minimizing protein aggregation. Amino acid combinations, glycine with glutamic acid, provided maximum stability. Formulation F4 (containing glycine, glutamic acid and Tween 20) showed minimum aggregation and degradation and in vivo activity equivalent to commercially available HSA-stabilized EPO (Eprex®).

Optimization of amino acid-stabilized erythropoietin parenteral formulation: <italic>In vitro</italic> and <italic>in vivo</italic> assessment

National Research Center, Dokki, Cairo 12311, Egypt

Department of Pharmaceutics College of Pharmacy, King Saud University, P.O. Box 2457, Riyadh 11451, Saudi Arabia

Pharmaceutical Sciences Department College of Pharmacy-3163, King Saud bin Abdulaziz University for Health Sciences, and King Abdullah International Medical, Research Center, Ministry of National, Guard, Health Affairs, Riyadh 11481, Saudi Arabia

Department of Pharmaceutics, Faculty of Pharmacy, Al-Azhar University, Cairo, Egypt

Acta Pharmaceutica

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Optimization of amino acid-stabilized erythropoietin parenteral formulation:  and  assessment

The aim of this study was to optimize the formulation of erythropoietin (EPO) using amino acids instead of human serum albumin (HSA) and to evaluate its in...

Optimization of amino acid-stabilized erythropoietin parenteral formulation: In vitro and in vivo assessment

Published Online: Mar 07, 2016

Page range: 69 - 82

Accepted: Sep 28, 2015

DOI: https://doi.org/10.1515/acph-2016-0007

Keywords
erythropoietin (EPO), amino acids, human serum albumin, protein stability, Bradford, ELISA, gel electrophoresis

© 2016 Bahgat E. Fayed et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Optimization of amino acid-stabilized erythropoietin parenteral formulation: In vitro and in vivo assessment

Published Online: Mar 07, 2016

Page range: 69 - 82

Accepted: Sep 28, 2015

DOI: https://doi.org/10.1515/acph-2016-0007

Keywordserythropoietin (EPO), amino acids, human serum albumin, protein stability, Bradford, ELISA, gel electrophoresis

© 2016 Bahgat E. Fayed et al., published by De Gruyter Open

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Keywords
erythropoietin (EPO), amino acids, human serum albumin, protein stability, Bradford, ELISA, gel electrophoresis