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All Journal HAYATI Journal of Biosciences Microbiology Indonesia Health Science Journal of Indonesia Medical Journal of Indonesia Indonesian Journal of Cancer The Indonesian Biomedical Journal Universa Medicina Jurnal Farmasi Galenika (Galenika Journal of Pharmacy) Molecular and Cellular Biomedical Sciences (MCBS) Spektra: Jurnal Fisika dan Aplikasinya Acta Biochimica Indonesiana JFIOnline Majalah Kesehatan Pharmamedika The New Ropanasuri Journal of Surgery Narra J Indonesian Journal of Cancer Makara Journal of Science Jurnal Kedokteran Meditek Proceedings Book of International Conference and Exhibition on The Indonesian Medical Education and Research Institute
Septelia I. Wanandi
Department Of Biochemistry And Molecular Biology, Faculty Of Medicine, Universitas Indonesia
Aerospace Engineering Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemistry Dentistry Earth & Planetary Sciences Electrical & Electronics Engineering Energy Health Professions Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Neuroscience Nursing Physics Public Health

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Expression and Purification of Novel Peptide-Based Recombinant Proteins Targeting Breast Cancer Stem Cells Agusta, Istiqomah; Triwidyaningtyas, Silvia; Bela, Budiman; Mardiana, Mardiana; Wanandi, Septelia Inawati
Proceedings Book of International Conference and Exhibition on The Indonesian Medical Education Research Institute Vol. 9 No. - (2025): Proceedings Book of International Conference and Exhibition on The Indonesian M
Publisher : Writing Center IMERI FMUI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.69951/proceedingsbookoficeonimeri.v9i-.327

Abstract

Introduction: Peptides with specific affinity toward breast cancer stem cells (BCSCs), including GYSASRSTIPGK and GAIRIRLSEPLS, have gained interest in targeted cancer diagnostics; however, the low abundance of BCSCs often results in limited detection sensitivity. This study addresses this limitation by conjugating BCSC-specific peptides to self-assembling Human Papillomavirus (HPV16) L1 protein, resulting in virus-like particles composed of 72 pentamers that provide substantial signal amplification. In addition, the SpyTag–SpyCatcher system derived from the Streptococcus pyogenes CnaB2 domain provides a robust strategy to enhance peptide–protein stability. Objectives: This study aimed to express and purify three recombinant proteins as key components for assembling a BCSC-specific diagnostic construct: HPV16 L1–SpyTag, SpyCatcher–GYSASRSTIPGK, and SpyCatcher–GAIRIRLSEPLS. Methods: Recombinant genes encoding the target proteins were expressed in Escherichia coli (E. coli) BL21(DE3) following IPTG induction for 1–4 hours. Bacterial cells were harvested and lysed under native or denaturing conditions, and recombinant proteins were purified using Ni-NTA affinity chromatography. Results: SDS–PAGE analysis showed clear bands at the expected molecular weights for HPV16 L1–SpyTag (58.5 kDa), SpyCatcher–GYSASRSTIPGK, and SpyCatcher–GAIRIRLSEPLS (~15 kDa), with induction-dependent increases in band intensity. The SpyCatcher–peptide fusions were predominantly soluble and purified under native conditions, whereas HPV16 L1–SpyTag was largely insoluble and required denaturing purification. Conclusion: HPV16 L1–SpyTag and SpyCatcher–peptide fusions were successfully expressed and purified in E. coli, establishing a versatile platform with potential to enhance BCSC-targeted detection and treatment strategies in breast cancer. 
Co-Authors . ANDI Abdul Halim Sadikin Adang Bachtiar Agnes Kurniawan Agusta, Istiqomah Alice Tamara Amalia, Ginva Amarila Malik Andi A. Victor ANDI YEHUDA ANDREW ROBERT COSSINS Ani R. Prijanti Ani Retno Prijanti Anto Sulaksono Asmarinah Ay Ly Margaret Ay Ly Margareth Ayu Suraduhita Bagus Ramasha Amangku Bethy S. Hernowo Budiman Bela CENMI MULYANTO Chatarina Umbul Wahyuni Daniel Marcellius Simadibrata Darmiati, Sawitri DARYL ROBERT WILLIAMS Dewi Muliaty Dewi Sumaryani Soemarko Dewi, Yulia Ratna Dimas Priantono Din Syafruddin Dwi Retna Lestari Elfahmi, Khalida Ikhlasiya Tajdar Gefariena Elisna Syahruddin Elvira Hinsch Elvira Yunita Erlia Anggrainy Sianipar Erlina, Linda Eziefule, Oluebube Magnificient Fadilah Fadilah Febriana C. Iswanti Febriana Catur Iswanti FERY ARDIANSYAH Fitriyadi Kusuma Franciscus D. Suyatna Frans Ferdinal Ghanny, Niken Rahmah H. J. Freisleben Habibah Muhiddin HANS-JOACHIM FREISLEBEN HANS-JOACHIM FREISLEBEN HANS-JOACHIM FREISLEBEN Harahap, Alida R. HARALD HUBER HARALD HUBER Hendrik Kurniawan Herbowo A. Soetomenggolo Ida Gunawan IMAN SANTOSO IMAN SANTOSO Irawan Yusuf Irene Dian Jörg Gromoll Jeanne A. Pawitan Joedo Prihartono Karina Teja Putri Kartolo, Wiwi A. Kawengian, Kevin Johanes Kevin Johanes Kawengian Lisnawati Mahdaleny Mahdaleny Mardiana Mardiana Maria Dara Novi Handayani, Maria Dara Novi Maria Maghdalena Melva Louisa MICHAEL THOMM Mizuno, Seiya Mohamad Sadikin Muchlis Ramli Muchtaruddin Mansyur Muhammad Arif Muhammad I. Saleh Muhammad Ilyas Mukharjon Mukharjon Murdani Abdullah Nabillah, Deya Adiby Nafrialdi Nafrialdi Nihayah, Silviatun Noorwati Sutandyo Noorwati Sutandyo Novi S. Hardiany Novi Silvia Hardiany Noza Hilbertina Nukman H. Moeloek Nukman Moeloek Nurjati C. Siregar Nurjati Chairani Siregar Paramita, Rafika I. Paramita, Rafika Indah Parwati Abadi Soekarno Puji B.S. Asih Purnamawati Purnamawati Purnomo Soeharso Puspita E. Wuyung Puspita Eka Wuyung Putri, Putu Indah Paramita Adi Raafqi Ranasasmita Radiana D. Antarianto Remizar Alpaddli Reni Paramita Resda Akhra Syahrani Resda Akhra Syahrani Rianto Setiabudy Rini Puspitaningrum Rini Puspitaningtum RONDANG ROEMIATI SOEGIANTO RONDANG ROEMIATI SOEGIANTO RONDANG ROEMIATI SOEGIANTO Rosari Saleh Rostika Flora Sari, Dewi H. Sarwono Waspadji Sekar Arumsari SEKAR ARUMSARI, SEKAR Sekarutami, Sri Mutya SERUNI K.U. FREISLEBEN SERUNI KUSUMA UDYANINGSIH FREISLEBEN Sianipar, Erlia Anggrainy Siregar, Nurjati Siufui Hendrawan Sonar Panigoro SRI HANDAYANI Sri S. Ningsih, Sri S. Sri S. Sudardjat Sri W. Dwintasari Sri W.A. Jusman Sri Widia A.Jusman Syahrani, Resda A. Syahrani, Resda Akhra Syaiful Ichwan Syarifah Dewi Syarifah Salma Tjahjono D. Gondhowiardjo Trifonia Pingkan Triwidyaningtyas, Silvia Vanny Narita Wahyuning Ramelan Warsito Purwo Taruno Watanabe, Yukihide Wawaimuli Arozal Wawan Mulyawan Wawan Mulyawan Wibisana, I GNG. Yalista Fatia Nadia Yefta Moenadjat Yohana Yohana Yohanna Sorontou Yuni Ahda Yurika Sandra ZESSINDA LUTHFA ZESSINDA LUTHFA