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en Recombinant Expression and Purification of Extracellular Domain of the Programmed CellDeath Protein Receptor زیست شناسی ملکولی Molecular Biology مقالات اصلی Original Article <div style="text-align: justify;">Background: The programmed cell deathprotein 1 (PD-1), which is a member of the CD28 receptor family, can negatively regulate antitumor immune responses by interacting with its ligands, PD-L1 or PD-L2. The PD-1–PD-L1 signaling pathway is a checkpoint mechanism that plays essential roles in downregulating immune responses in cancerous tissues. Thus, blocking this signaling pathway leads to enhanced antitumor immunity, potentially preventing tumor progression. Methods: We synthesized the extracellular domain of the PD-1 receptor (rPD-1) de novoby using a two-step polymerase chain reaction and the Phusion® DNA polymerase. The synthesized gene was cloned into the pET28 expression plasmid and transformed into competent Escherichia coli. Purification of rPD-1 was performed by metal-affinity chromatography, using a HisTrap column.Purified rPD-1 was characterized by western blotting and mass spectrometry using the SwissProt database and the Mascot program. Results: Designed and synthesized construct of rPD-1 was 500 bpin size. Analysis of the electrophoresis data of purified rPD-1 showed the presence of a protein with a molecular mass of 21 kDa. Mass spectrometry data using the SwissProt database and the Mascot program outputted the highest-scoring sequence to correspond to rPD-1. Conclusions: Synthesized de novo rPD-1 may have potential therapeutic applications in enhancing antitumor immune responses</div> Cancer, PD-1 ligands, PD-1 receptor, Tumor immunotherapy 347 357 http://rbmb.net/browse.php?a_code=A-10-350-1&slc_lang=en&sid=1 Adish Zhansaya zhansaya.adish@gmail.com 100319475328460017392 100319475328460017392 Yes National Center for Biotechnology,Kurgalzhyn road, 13/5, Astana, 010000, Kazakhstan & L.N.GumilyovEurasian National University, Satpayev st., 2, Astana, 010008, Kazakhstan. Mukantayev Kanatbek mukantaev@biocenter.kz 100319475328460017393 100319475328460017393 No National Center for Biotechnology,Kurgalzhyn road, 13/5, Astana, 010000, Kazakhstan Tursunov Kanat kanat_tka@mail.ru 100319475328460017394 100319475328460017394 No National Center for Biotechnology,Kurgalzhyn road, 13/5, Astana, 010000, Kazakhstan Ingirbay Bakhytkali bakytkali.kabi@mail.ru 100319475328460017395 100319475328460017395 No National Center for Biotechnology,Kurgalzhyn road, 13/5, Astana, 010000, Kazakhstan Kanayev Darkhan dake_2008z@mail.ru 100319475328460017396 100319475328460017396 No National Center for Biotechnology,Kurgalzhyn road, 13/5, Astana, 010000, Kazakhstan Kulyyassov Arman akulyyasov@gmail.com 100319475328460017397 100319475328460017397 No National Center for Biotechnology,Kurgalzhyn road, 13/5, Astana, 010000, Kazakhstan Tarlykov Pavel tarlykov@biocenter.kz 100319475328460017398 100319475328460017398 No National Center for Biotechnology,Kurgalzhyn road, 13/5, Astana, 010000, Kazakhstan Mukanov Kasym mukanov@biocenter.kz 100319475328460017399 100319475328460017399 No National Center for Biotechnology,Kurgalzhyn road, 13/5, Astana, 010000, Kazakhstan Ramankulov Yerlan ramanculov@biocenter.kz 100319475328460017400 100319475328460017400 No National Center for Biotechnology,Kurgalzhyn road, 13/5, Astana, 010000, Kazakhstan