Melola Discovery

← Melola Discovery

Akses terbuka Jurnal SINTA 1

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) · hlm. 51–58

DOI
10.11594/jtls.15.01.06
ISSN
2527-4376, 2087-5517
Terbit
2025-06-26
Penerbit
Journal of Tropical Life Science
Bahasa
eng
Lisensi
creativecommons.org/licenses/by-nc/4.0

Abstrak

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.

Kata kunci

Jaringan kolaborasi para penulis

40 penulis · 157 tautan kepenulisan bersama · 2 lintas organisasi · menampilkan 40 yang paling banyak terhubung

Arahkan kursor atau ketuk sebuah nama untuk menyorot rekan penulisnya. Klik dua kali untuk membuka halaman penulis.

  • University of Brawijaya
  • Hasanuddin University
  • Afiliasi tidak diketahui
  • Organisasi berbeda
  • Organisasi sama
  • Afiliasi tidak diketahui
  • Garis lebih tebal = lebih banyak artikel bersama · lingkaran lebih besar = lebih banyak artikel
Tampilkan sebagai tabel
Rekan penulisOrganisasiArtikel bersama
Sri RahayuHasanuddin University6
Aris SoewondoAfiliasi tidak diketahui5
Sri WidyartiAfiliasi tidak diketahui5
Fikriya Novita SariAfiliasi tidak diketahui4
Farida RachmawatiAfiliasi tidak diketahui3
Sefihara Paramitha SaktiAfiliasi tidak diketahui3
Dinia Rizqi DwijayantiAfiliasi tidak diketahui3
Moh DliyauddinAfiliasi tidak diketahui2
Hideo TsuboiAfiliasi tidak diketahui2
Tri ArdyatiAfiliasi tidak diketahui2
Irfan MustafaAfiliasi tidak diketahui2
Muhammad Halim NatsirUniversity of Brawijaya2
Mansur IbrahimAfiliasi tidak diketahui2
Rahmat GrahadiAfiliasi tidak diketahui2
Muhammad Hermawan WidyanandaAfiliasi tidak diketahui2
Wayan Firdaus MahmudyUniversity of Brawijaya2
Achmad RidokUniversity of Brawijaya2
Muhammad Sasmito DjatiAfiliasi tidak diketahui2
Sofia Nur CahyaniAfiliasi tidak diketahui1
Belinda Nabiila Al FaizahAfiliasi tidak diketahui1
Rif'atul Hawani. BAfiliasi tidak diketahui1
Nabila Shafa Yumna SalsabilaAfiliasi tidak diketahui1
Noviana Dwi LestariAfiliasi tidak diketahui1
Agung Pramana Warih MahendraAfiliasi tidak diketahui1
Agung Putra PurwantoAfiliasi tidak diketahui1
Alifia Issabella MulyawatiAfiliasi tidak diketahui1
Elsa Mega SuryaniAfiliasi tidak diketahui1
Hikmah MaulidiyahAfiliasi tidak diketahui1
Salam A. IbrahimAfiliasi tidak diketahui1
Firda Nuri AsyhariAfiliasi tidak diketahui1
Heni Sukma ZulfatimAfiliasi tidak diketahui1
Nenis Try Melani PutriAfiliasi tidak diketahui1
Ahmad Shobrun JamilAfiliasi tidak diketahui1
asep awaludin prihantoAfiliasi tidak diketahui1
Rika Gian MeliantiAfiliasi tidak diketahui1
Titiek Indhira AgustinAfiliasi tidak diketahui1

Afiliasi berasal dari OpenAlex; metadata OAI-PMH jurnal tidak memuatnya. Penulis tanpa ID OpenAlex dicocokkan berdasarkan nama, sehingga dua orang bisa tergabung atau satu orang bisa terpisah.

Sumber metadata: OAI-PMH (metadata jurnal) http://jtrolis.ub.ac.id/index.php/jtrolis/oai, pertama di-harvest 9 Nov 2026, terakhir terlihat 9 Nov 2026.
Metadata asli (oai_dc) 
<oai_dc:dc xmlns="http://www.openarchives.org/OAI/2.0/" xmlns:dc="http://purl.org/dc/elements/1.1/" xmlns:oai_dc="http://www.openarchives.org/OAI/2.0/oai_dc/" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xsi:schemaLocation="http://www.openarchives.org/OAI/2.0/oai_dc/  http://www.openarchives.org/OAI/2.0/oai_dc.xsd">
	<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>