Detection of Recombinant Erythropoietin Biosimilars and Analog Substances in Human Urine by Gel Electrophoresis Technique.
abdallah, A., Abo-El-Matty, D., Mesbah, N., Wahba, A., Ramadan, A. (2024). Detection of Recombinant Erythropoietin Biosimilars and Analog Substances in Human Urine by Gel Electrophoresis Technique.. EKB Journal Management System, 8(1), 169-178. doi: 10.21608/rpbs.2024.292878.1327
ahmed anas abdallah; Dina Abo-El-Matty; Noha Mesbah; Alaa Samir Wahba; Asmaa Ramadan. "Detection of Recombinant Erythropoietin Biosimilars and Analog Substances in Human Urine by Gel Electrophoresis Technique.". EKB Journal Management System, 8, 1, 2024, 169-178. doi: 10.21608/rpbs.2024.292878.1327
abdallah, A., Abo-El-Matty, D., Mesbah, N., Wahba, A., Ramadan, A. (2024). 'Detection of Recombinant Erythropoietin Biosimilars and Analog Substances in Human Urine by Gel Electrophoresis Technique.', EKB Journal Management System, 8(1), pp. 169-178. doi: 10.21608/rpbs.2024.292878.1327
abdallah, A., Abo-El-Matty, D., Mesbah, N., Wahba, A., Ramadan, A. Detection of Recombinant Erythropoietin Biosimilars and Analog Substances in Human Urine by Gel Electrophoresis Technique.. EKB Journal Management System, 2024; 8(1): 169-178. doi: 10.21608/rpbs.2024.292878.1327

Detection of Recombinant Erythropoietin Biosimilars and Analog Substances in Human Urine by Gel Electrophoresis Technique.

Records of Pharmaceutical and Biomedical Sciences
Volume 8, Issue 1, 2024, Page 169-178  XML PDF (680.85 K)
Document Type: Original Article
DOI: 10.21608/rpbs.2024.292878.1327
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Authors
ahmed anas abdallah email 1; Dina Abo-El-Mattyorcid 2; Noha Mesbahorcid 3; Alaa Samir Wahbaorcid 4; Asmaa Ramadan3
1Biochemistry, pharmacy, Suez canal university, Ismailia.
2Faculty of Pharmacy - Suez Canal University
3Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt.
4Department of Biochemistry, Faculty of Pharmacy, Suez Canal University, Ismailia, Egypt
Abstract
This study aimed to improve the detection of erythropoietin (EPO) and its biosimilars in human urine using the SAR-PAGE technique by electrophoresis method in pre-casted gels to help differentiate each biosimilar of EPO according to their molecular masses along with their electrophoretic characters. 20 ml human urine samples were collected, purified then spiked with the known concentration of erythropoietin standards. SAR-PAGE electrophoresis was applied using two different concentrations of N-Lauroylsarcosine sodium salt to visualize the difference and effect of sarkosyl on human erythropoietin detection, followed by immunodetection using chemiluminescence reagent. All data were collected, tabled, and statistically analyzed. The first group (using 0.1% w/v SARCOSYL-PAGE) showed large condensed shadowed bands and asymmetric bands behavior while the second group (using 0.12 % w/v SARCOSYL-PAGE) showed confirmed, thin,sharp bands and symmetric identical bands behavior. Increasing sarcosyl concentration to 0.12 % w/v affect the immunological detection by increasing the affinity of the monoclonal anti-EPO antibody to the protein chain in both PEGylated and non-PEGylated epoetins thus leading to enhanced antibody binding and a sharper electrophoretic separation and detectable bands.
Keywords
Doping; Mircera; SARCOSYL-PAGE; Immunopurification; Chemiluminescence
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