Production and characterization of long-acting recombinant human albumin–EPO fusion protein expressed in CHO cell

Joung, Chan-Hi; Shin, Ju-Yeop; Koo, Jae-Kyung; Lim, Jin J.; Wang, Jin-Sang; Lee, Song-Jae; Tan, Hyun-Kwang; Kim, Sang-Lin; Lim, Sang‐Min

doi:10.1016/j.pep.2009.07.003

Cited by 38 publications

(17 citation statements)

References 33 publications

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“…Binding occurs only under acidic pH-conditions as encountered within endosomes (pH 6-6.4), but not under physiological pH of 7.4. [8][9][10][11][12][13][14][15] All of these modifications decrease the affinity of EPO to its receptor (EPO-R) and delay clearance of EPO from the blood stream due to less efficient glomerular filtration. Erythropoietin (EPO), the protein hormone responsible for differentiation of bone marrow stem cells into erythrocytes, has been recombinantly produced since 1985 and was first approved for clinical application in anaemia treatment in 1989.…”

Section: Introductionmentioning

confidence: 99%

“…albumin, IgG-Fc, granulocyte/monocyte colony stimulating factor (GM-CSF), hCG). [8][9][10][11][12][13][14][15] All of these modifications decrease the affinity of EPO to its receptor (EPO-R) and delay clearance of EPO from the blood stream due to less efficient glomerular filtration. Fusion proteins consisting of EPO and the Fc-part of human IgG have the additional advantage of opening the route of pulmonary administration via the neonatal Fc receptor.…”

Section: Introductionmentioning

confidence: 99%

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Detection of EPO‐Fc fusion protein in human blood: Screening and confirmation protocols for sports drug testing

Reichel

Thevis

2012

Drug Testing and Analysis

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The neonatal Fc receptor (FcRn) has been under investigation for several years as a pharmaceutical drug target. Clinical studies have shown that fusion proteins consisting of human recombinant erythropoietin (rhEPO) and the Fc-part of IgG can be transported after pulmonary administration via FcRn across the airway epithelium to the blood stream. So far, no clinically approved pharmaceutical formulation of EPO-Fc is available. Since various forms of recombinant erythropoietins have been frequently misused by athletes as performance-enhancing agents, EPO-Fc might play a similar role in sports in the future. In order to investigate the detectability of EPO-Fc in human blood, different strategies were tested and developed. Only two of them fulfilled the necessary requirements regarding sensitivity and specificity. A rapid protocol useful for screening purposes first enriches EPO-Fc from human serum via high capacity protein A beads and subsequently detects EPO-Fc in the eluate with a commercial EPO ELISA kit. The limit of detection (LOD) of the method is about 5 pg (45 amol) EPO-Fc and is independent of the serum volume used. For screening and/or confirmation purposes a second protocol was evaluated, which consists of a fast EPO immunopurification step followed by sodium dodecyl sulfate or sarcosyl polyacrylamide gel electrophoresis (SDS-PAGE, SAR-PAGE) and Western double-blotting with chemiluminescence detection - a method already established in routine EPO anti-doping control. The latter strategy allows the detection of EPO-Fc in serum together with all other recombinant erythropoietins and with an identical LOD (5 pg/45 amol) as for the rapid screening protocol.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Detection of EPO‐Fc fusion protein in human blood: Screening and confirmation protocols for sports drug testing

Reichel

Thevis

2012

Drug Testing and Analysis

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“…30 Glycosylation can increase levels of terminal sialylation for HuEPO and improve protein half-life as well as resistance to proteolysis. 31,32 Therefore, glycosylation impacts HuEPO stability. 17,33 On the other hand, some studies proposed that N-glycans with sialic acid extensions are not the main factors in of HuEPO biological activity.…”

Section: Discussionmentioning

confidence: 99%

Aggregate Forms of Recombinant Human Erythropoietin With Different Charge Profile Substantially Impact Biological Activities

Ghezlou

Mokhtari

Kalbasi

et al. 2020

Journal of Pharmaceutical Sciences

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“…The binding and in vitro anti-viral activity of CD4 for the albumin-CD4 fusion was similar to those of soluble CD4 [227]. Further development of albumin fusion technologies has focused on proteins that have been used clinically, for example, cytokines including INF-b [48], INF-a2b (Albuferon) [47,228], growth hormone (Albutropin) [229], G-CSF (Albugranin) [46], EPO [230] and interleukin-2 (IL-2) (Albuleukin) [231]. Additional examples include hirudin [232], factor VIIa [233], and thioredoxin-1 (Trx) [234] and bispecific antibodies [235].…”

Section: Fusion Proteinsmentioning

confidence: 99%

Chemical and Genetic Modification

Farys¹,

Ginn²,

Badescu³

et al. 2013

Pharmaceutical Sciences Encyclopedia

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There are many natural post‐translational modifications of proteins (e.g., glycosylation, ubiquitination, acylation,and amino acid derivatization).Glycosylation is common for therapeutic proteins. Classes of proteins modified by glycosylation include monoclonal antibodies, blood factors, hematopoietic proteins, and cytokines. Excluding monoclonal antibodies, many proteins were, at least when they were first introduced to the clinic, designed to be used as a replacement protein for the corresponding endogenous protein. Maintaining control of the protein structure to be as close as possible to the structure of the endogeneous protein is therefore a key goal. In this review, we focus on (i) optimization of protein pharmacokinetics, (ii) improvement of protein properties to be used as medicines (this primarily includes reduction of immunogenic sequences and improvement of protein stability), (iii) addition of a therapeutic effect, and (iv) use of proteins as diagnostics.

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Production and characterization of long-acting recombinant human albumin–EPO fusion protein expressed in CHO cell

Cited by 38 publications

References 33 publications

Detection of EPO‐Fc fusion protein in human blood: Screening and confirmation protocols for sports drug testing

Detection of EPO‐Fc fusion protein in human blood: Screening and confirmation protocols for sports drug testing

Aggregate Forms of Recombinant Human Erythropoietin With Different Charge Profile Substantially Impact Biological Activities

Chemical and Genetic Modification

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