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All Journal AGRITROP Jurnal Ilmu Dasar Melintas An International Journal of Philosophy and Religion Prosiding Seminar Biologi JURNAL TEKNOLOGI LINGKUNGAN AL KAUNIYAH BERKALA SAINSTEK Biota: Jurnal Ilmiah Ilmu-Ilmu Hayati Industrial and Systems Engineering Assessment Journal (INASEA) Agrointek Indonesian Journal of Agricultural Science Jurnal AgroBiogen Buletin Penelitian Tanaman Rempah dan Obat AGRIVITA, Journal of Agricultural Science ROTOR: JURNAL ILMIAH TEKNIK MESIN UNEJ e-Proceeding Extension Course Filsafat ( ECF ) Agriprima : Journal of Applied Agricultural Sciences Jurnal Bioteknologi & Biosains Indonesia (JBBI) Jurnal Penelitian Pendidikan IPA (JPPIPA) Jurnal ASET (Akuntansi Riset) ANNALES BOGORIENSES Mesin Agritrop : Jurnal Ilmu-Ilmu Pertanian (Journal of Agricultural Science) Jurnal Pengabdian Magister Pendidikan IPA YUME : Journal of Management ACCRUALS (Accounting Research Journal of Sutaatmadja) Jurnal Permukiman J-Dinamika: Jurnal Pengabdian Kepada Masyarakat JURNAL REFORMASI ADMINISTRASI Jurnal Ilmiah untuk Mewujudkan Masyarakat Madani Journal Of Management Science (JMAS) Jurnal Review Pendidikan dan Pengajaran (JRPP) Jurnal Mantik Journal of Contemporary Accounting Jurnal Informasi dan Teknologi Jurnal JTIK (Jurnal Teknologi Informasi dan Komunikasi) JASS (Journal of Accounting for Sustainable Society) Buletin Penelitian Tanaman Rempah dan Obat Economics and Digital Business Review TSARWATICA (Islamic Economic, Accounting, and Management Journal) Operations Research: International Conference Series Jurnal Pengabdian Masyarakat (ABDIRA) International Journal of Quantitative Research and Modeling Proceeding Biology Education Conference Golden Ratio of Marketing and Applied Psychology of Business BIOVALENTIA: Biological Research Journal Golden Ratio of Mapping Idea and Literature Format Golden Ratio of Auditing Research Journal Of Human And Education (JAHE) Golden Ratio of Data in Summary Journal of Artificial Intelligence and Digital Business Journal of Law, Education and Business Kajian Akuntansi Jurnal Medika: Medika Biosaintifika: Journal of Biology & Biology Education
Bambang Sugiharto
Fakultas Matematika Dan Ilmu Pengetahuan Alam Universitas Jember
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Journal : AL KAUNIYAH

 1 
Produksi Antibodi Poliklonal Menggunakan Protein Rekombinan RBD-spike Untuk Deteksi SARS-CoV-2 Iryani Endah Febrianti; Yayuk Fatmawati; Intan Ria Neliana; Widhi Dyah Sawitri; Erlia Narulita; Bambang Sugiharto
Al-Kauniyah: Jurnal Biologi Vol 16, No 2 (2023): AL-KAUNIYAH JURNAL BIOLOGI
Publisher : Department of Biology, Faculty of Science and Technology, Syarif Hidayatullah State Islami

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15408/kauniyah.v16i2.25664

Abstract

 AbstrakSARS-CoV-2 merupakan virus yang menyebabkan Coronavirus Disease 2019 (COVID-19) di seluruh dunia dan sampai saat ini kasus terbaru masih terus dilaporkan. Diagnostic test merupakan hal yang krusial untuk dikembangkan. Prinsip diagnostic test COVID-19 berbasis antigen, yaitu mendeteksi virus SARS-CoV-2 melalui respon antibodi dari penderita. Penelitian ini bertujuan untuk memproduksi antibodi poliklonal menggunakan protein rekombinan RBD-Spike untuk mendeteksi virus SARS-CoV-2 berbasis antibodi. Penelitian dimulai dengan penentuan domain RBD-Spike menggunakan pensejajaran asam amino, dan konstruksi DNA untuk RBD-Spike pada vektor ekspresi pET28a menggunakan sintetik nukleotida. Produksi protein rekombinan RBD-Spike diekspresikan pada sel bakteri Escherichia coli. Purifikasi dilakukan untuk memperoleh protein RBD-Spike dan selanjutnya digunakan sebagai antigen untuk induksi antibodi poliklonal pada kelinci. Hasil penelitian menunjukkan bahwa ekspresi protein rekombinan RBD-Spike SARS-CoV-2 memerlukan induksi IPTG 0,1 mM dan terekspresi dalam bentuk inclusion bodies dengan ukuran 39 kDa. Purifikasi protein RBD-Spike dilakukan menggunaan resin afinitas NiNTA, elektroelusi, dan dialisis. Total protein RBD-Spike yang diperoleh sebanyak 4 mL dengan konsentrasi 10 mg/mL. Analisa Ouchterlony menunjukkan bahwa antibodi poliklonal terdeteksi pada minggu kedua setelah injeksi booster dan analisa spesifitas antibodi terhadap antigen menunjukkan bahwa antibodi poliklonal dapat mendeteksi protein RBD-Spike pada konsentrasi 0,1 µg. Selanjutnya diharapkan antibodi poliklonal dapat digunakan untuk deteksi keberadaan virus SARS-CoV-2 dan dapat dikembangkan untuk kit deteksi berbasis antibodi.AbstractSARS-CoV-2 is the virus that causes Coronavirus Disease 2019 (COVID-19) worldwide and the latest cases are still being reported until now. The diagnostic test is a crucial to be developed. The principle of the antigen-based COVID-19 diagnostic test is to detect the SARS-CoV-2 virus through antibody response from the patients. This study was conducted to produce polyclonal antibodies using recombinant protein RBD-Spike. The research was carried out by determining the RBD-Spike domain using amino acid alignment and constructing the DNA of RBD-Spike to the expression vector of pET28a using nucleotide synthesis. Production of RBD-Spike recombinant protein was expressed in Escherichia coli. Purification was carried out to obtain RBD-Spike protein and used to induce polyclonal antibody ina rabbit. The results showed that the expression of RBD-Spike recombinant protein required induction of IPTG 0.1 mM and was expressed in inclusion bodies with molecular size of 39 kDa. The purification of RBD-Spike protein was carried out using resin affinity, electroelution, and dialysis. The total protein of RBD-Spike obtained was 4 mL with a concentration of 10 mg/mL. Ouchterlony analysis revealed that polyclonal antibody was detected in the second week after booster injection and analysis of antibody specificity showed that polyclonal antibodies detected RBD-Spike protein at the concentration of 0.1µg of RBD-Spike protein. Moreover, it is expected that our polyclonal antibody detect the SARS-CoV-2 virus and can be developed for antibody-based detection kits.
Co-Authors A Bagus Nur Sudrajat A. Dinar Ambarwati Agung Nur Probohudono Agus Marwan Agustien, Inyana Dwi Agustien, Inyana Dwi Ahmad Hidayat Ahmad Ilham Tanzil Ai Sumirah Setiawati Amir, Muhammad Hisyam Andriani, Maya Anita Firmanti Anjas Wida Elistia Rini Anuary Dwi Rosyid, Wafa Prasetia Arif Syaifudin, Arif Ariyono, Alief Rizky Asshidiqie, Wahyu Pratama Aventi Aventi, Aventi Azizah, Kunti Anis Bambang Subiyanto, Bambang Bambang Sugiharto Barus, Muhammad Arif Bayati, Tutiana Bon, Abdul Thalib Bin Budi Sulistiarini, Emma Dany Cahyadi De Hans, Nickytha Linch Dedi Dwilaksana Dedy Eko Rahmanto Dewita, Tiara Ajeng Dian Al Ghifari Perwitasari Digdo Listyadi, Digdo Djenal . Dr. Hadrianus Tedjoworo, S.Ag., STL. Dwi Djumhariyanto Edia F.D Edia, Edia F.D Edison Edison Edy Listanto Elizabeth Eneng Tita Tosida Eny I. Riyanti Erlia Narulita Esti Utarti Fajrianto, Sigit Fanata, Wahyu Indra Duwi Fatimah Zahara Fibriani, Suwinda Gaol, Nickytha Linch De Hans Lumban Gulo, Desi Natalia Hafiz, Syauqi Abyan Hafizi, Muhamad Halawa, Herry Martinus Hari Purnomo HARKIM Harkim, H. Harkim, Harkim Hasibuan, Aqil Muhammad Hayati, Nur Fadillah Hiya, Nirmadarningsih Husin, Andriati Amir Icih, Icih Ika Oktavianawati Ilhamsyah Siregar Imas Cintamulya Insani, Vivian Nurul Intan Ria Neliana Inyana Dwi Agustien Irawan Gulo, Sastra Darma Iryani Endah Febrianti Jayus Jayus Kesuma, Muhammad Yudha Kurnia Indriyanti Purnamasari, Kurnia Indriyanti Kurniawan , Asep Kusmana Kusmana Laily Ilman Widuri, Laily Ilman Laksono Trisnantoro Lasino Lasino, Lasino Lassa, Anita Lathifah Hanum, Lathifah Liem, Jhon Sepron Defretus Liliek Sulistyowati Lilik Yulianingsih Lina Maulidiana Maesaroh, Nurlaila Magnon, Cassidy Maisaro, Maisaro Marina Pakaya Marwan, Agus Maulani, Ardela Alief Maulida, Dinda Mala MAYA ANDRIANI Media Ningtyas, Rinda MISWAR - Mochamad Asrofi Mohd Radzi, Fathin Nuril A'isha Binti Muhammad Hazmi Muhammad, Adibillah Mukhamad Su'udi Nadya Oktafiana Neliana, Intan Ria Netty Ermawati Nikmah, Nasilatun Nina Oktaria, Nina Ningtiyas, Wulan Nursyiam Nirmadarningsih Hiya Nirwana Br Bangun Nugroho, Sandi Luqman Nur Basuki Nurfatimah, Siti Nurhayati Nursely, Dinda Oktapan, Ladefa Primana Pandiangan, Jenny Ester Parawita Dewanti Purnama Okviandari, Purnama Purwinto, Rahmanaji Setyo Putri, Trisandi Eka Rachman Setiawan Rahardhianto, Arsetyo Raharja, Ajung Gilang Putra Raharjo, Sulistyo Yusuf Rahma Rei Sakura Ramadhan, Mohamad Ibnu Ramadhani, Rifki Fahrisal Ratnasari, Tri Ratri Ciptaningtyas Rejekia V Simanungkalit Restanto, Didik Pudji Rizal, Asep Rizki, Ainur Salahuddin Junus Salih, Yasir Sari , Indaniaty Hasanah Sigit Soepardjono Simamora, Harkim Simanungkalit, Rejekia Simanungkalit, Rejekia V Simanungkalit, Rejekia V. Simanungkalit, Rejekia Vaizal Sinambela, Khayriza Sirait, Syah Paradiba Siregar, Ilhamsyah Siregar, Luthfi Hidayat Siti Arifah Siti Rohimah Siti Umayyah Slameto Slameto Soleh, Mohamad Badrus SRI AYU MADE Sri Mulyati Sri Mulyati Sri Panca Setyawati Sri Rejeki Subandi, Kotim Sugara, Victor Ilyas Sugianto, Efendi Sugiarto, Aan Sukardiman Syafina Pusparani Tanjung, Yulia Tiara Toto Hadiarto Tri Agus Siswoyo TRI HANDOYO Tri J. Santoso Tri Ratnasari Ummi Sholikhah Untung Murdiyanto Wahyu Nurkholis Hadi Syahputra Widhi Dyah Sawitri Widhiya Ninsiana, Widhiya Wulan Nursyiam Ningtyas Yanti Setianti Yayuk Fatmawati YENNI SAMRI JULIATI NASUTION, YENNI SAMRI JULIATI Yoga Yuniadi Yudi Rinanto Yundari, Yundari Yusuf Rachmandika