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Effect of Combination of Different Antibiotics and Promoters for Expressing Recombinant Darbepoetin in Stable CHO K-1 Cell Line: Evaluating Antibiotic Combinations to Improve Yield and Quality of Darbepoetin

Journal of Tropical Life Science : International Journal of Theoretical, Experimental, and Applied Life Sciences · Vol. 15 No. 1 (2025) · pp. 51–58

DOI
10.11594/jtls.15.01.06
ISSN
2527-4376, 2087-5517
Published
2025-06-26
Publisher
Journal of Tropical Life Science
Language
eng
License
creativecommons.org/licenses/by-nc/4.0

Abstract

Antibiotics are key for successful molecular cloning techniques. Different antibiotics have different mechanisms of action, which leads to cell heath and a viable number of passages. Moreover, the suitability of the promoter also plays an important role in achieving a higher level of protein titter in the stable cell platform. Therefore, with plenty of options of antibiotics and promoters available, we need to determine the best combination of antibiotics and promoters, particularly for specific proteins of interest. Darbepoetin is a recombinant therapeutics protein with extra glycosylation to increase the half-life in the blood; this drug is used for the administration of CKD and leukemia patients. Blasticidin-S and puromycin were used as antibiotics, and CMV and EF-1 promoters were used in this experiment to evaluate the expression of recombinant darbepoetin for the protein model. CHO K-1 cell line was transfected with a plasmid carrying a combination of promoter and antibiotics genes; after 14 days, the level of specific protein expression was evaluated using the western blot technique. A single clone cell was obtained using the serial dilution method to evaluate the clonality and expression of the protein of interest. This study successfully obtained a single clone from stable pool transfection. This result suggested that a combination of puromycin antibiotics and EF-1 promoter has promising expression compared to Blasticidin-S antibiotics with CMV promoter. For further conclusion, an analytical comparison of both combinations needed to be done.

Keywords

Collaboration network of the authors

40 authors · 157 co-authorship links · 2 across different organisations · showing the 40 most connected

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  • University of Brawijaya
  • Hasanuddin University
  • Affiliation unknown
  • Different organisations
  • Same organisation
  • Affiliation unknown
  • Thicker line = more joint articles · bigger circle = more articles
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Co-authorOrganisationJoint articles
Sri RahayuHasanuddin University6
Aris SoewondoAffiliation unknown5
Sri WidyartiAffiliation unknown5
Fikriya Novita SariAffiliation unknown4
Farida RachmawatiAffiliation unknown3
Sefihara Paramitha SaktiAffiliation unknown3
Dinia Rizqi DwijayantiAffiliation unknown3
Moh DliyauddinAffiliation unknown2
Hideo TsuboiAffiliation unknown2
Tri ArdyatiAffiliation unknown2
Irfan MustafaAffiliation unknown2
Muhammad Halim NatsirUniversity of Brawijaya2
Mansur IbrahimAffiliation unknown2
Rahmat GrahadiAffiliation unknown2
Muhammad Hermawan WidyanandaAffiliation unknown2
Wayan Firdaus MahmudyUniversity of Brawijaya2
Achmad RidokUniversity of Brawijaya2
Muhammad Sasmito DjatiAffiliation unknown2
Sofia Nur CahyaniAffiliation unknown1
Belinda Nabiila Al FaizahAffiliation unknown1
Rif'atul Hawani. BAffiliation unknown1
Nabila Shafa Yumna SalsabilaAffiliation unknown1
Noviana Dwi LestariAffiliation unknown1
Agung Pramana Warih MahendraAffiliation unknown1
Agung Putra PurwantoAffiliation unknown1
Alifia Issabella MulyawatiAffiliation unknown1
Elsa Mega SuryaniAffiliation unknown1
Hikmah MaulidiyahAffiliation unknown1
Salam A. IbrahimAffiliation unknown1
Firda Nuri AsyhariAffiliation unknown1
Heni Sukma ZulfatimAffiliation unknown1
Nenis Try Melani PutriAffiliation unknown1
Ahmad Shobrun JamilAffiliation unknown1
asep awaludin prihantoAffiliation unknown1
Rika Gian MeliantiAffiliation unknown1
Titiek Indhira AgustinAffiliation unknown1

Affiliations come from OpenAlex; journal OAI-PMH metadata carries none. Authors without an OpenAlex ID are matched by name, which can merge or split people.

Metadata source: OAI-PMH (metadata jurnal) http://jtrolis.ub.ac.id/index.php/jtrolis/oai, first harvested 9 Nov 2026, last seen 9 Nov 2026.
Original metadata (oai_dc) 
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	<dc:title xml:lang="en">Effect of Combination of Different Antibiotics and Promoters for Expressing Recombinant Darbepoetin in Stable CHO K-1 Cell Line: Evaluating Antibiotic Combinations to Improve Yield and Quality of Darbepoetin</dc:title>
	<dc:creator xml:lang="en">Widekdo, Dwi purno</dc:creator>
	<dc:creator xml:lang="en">Widodo, Nashi</dc:creator>
	<dc:creator xml:lang="en">Rifa'i, Muhaimin</dc:creator>
	<dc:creator xml:lang="en">Jatmiko, Yoga Dwi</dc:creator>
	<dc:subject xml:lang="en">Antibiotics resistance</dc:subject>
	<dc:subject xml:lang="en">Clone selection</dc:subject>
	<dc:subject xml:lang="en">Promoter</dc:subject>
	<dc:subject xml:lang="en">protein recombinant</dc:subject>
	<dc:subject xml:lang="en">protein expression</dc:subject>
	<dc:description xml:lang="en">Antibiotics are key for successful molecular cloning techniques. Different antibiotics have different mechanisms of action, which leads to cell heath and a viable number of passages. Moreover, the suitability of the promoter also plays an important role in achieving a higher level of protein titter in the stable cell platform. Therefore, with plenty of options of antibiotics and promoters available, we need to determine the best combination of antibiotics and promoters, particularly for specific proteins of interest. Darbepoetin is a recombinant therapeutics protein with extra glycosylation to increase the half-life in the blood; this drug is used for the administration of CKD and leukemia patients. Blasticidin-S and puromycin were used as antibiotics, and CMV and EF-1 promoters were used in this experiment to evaluate the expression of recombinant darbepoetin for the protein model. CHO K-1 cell line was transfected with a plasmid carrying a combination of promoter and antibiotics genes; after 14 days, the level of specific protein expression was evaluated using the western blot technique. A single clone cell was obtained using the serial dilution method to evaluate the clonality and expression of the protein of interest. This study successfully obtained a single clone from stable pool transfection. This result suggested that a combination of puromycin antibiotics and EF-1 promoter has promising expression compared to Blasticidin-S antibiotics with CMV promoter. For further conclusion, an analytical comparison of both combinations needed to be done.</dc:description>
	<dc:publisher xml:lang="en">Journal of Tropical Life Science</dc:publisher>
	<dc:date>2025-06-26</dc:date>
	<dc:type>info:eu-repo/semantics/article</dc:type>
	<dc:type>info:eu-repo/semantics/publishedVersion</dc:type>
	<dc:format>application/pdf</dc:format>
	<dc:identifier>https://jtrolis.ub.ac.id/index.php/jtrolis/article/view/3373</dc:identifier>
	<dc:identifier>10.11594/jtls.15.01.06</dc:identifier>
	<dc:source xml:lang="en">Journal of Tropical Life Science; Vol. 15 No. 1 (2025); 51-58</dc:source>
	<dc:source>2527-4376</dc:source>
	<dc:source>2087-5517</dc:source>
	<dc:language>eng</dc:language>
	<dc:relation>https://jtrolis.ub.ac.id/index.php/jtrolis/article/view/3373/779</dc:relation>
	<dc:rights xml:lang="en">Copyright (c) 2025 Journal of Tropical Life Science</dc:rights>
	<dc:rights xml:lang="en">https://creativecommons.org/licenses/by-nc/4.0</dc:rights>
</oai_dc:dc>