ORIGINAL_ARTICLE Introduction: Endotoxin removal is a crucial stage in ensuring the safety of parenteral products. S3E3-S-Sepharose, which has been generated via site-specific immobilization of the S3E3 cationic amphiphilic peptide (CAP) on Sepharose, is a newly designed affinity matrix proposed for lipopolysaccharide (LPS) removal from biopharmaceuticals and vaccines. Methods: In the current study, the kinetic behavior of LPS adsorption on the matrix was investigated at pHs 4.5 and 8.5 by incubation of LPS-contaminated bovine serum albumin (BSA) solutions, as a model, with the S3E3-Sepharose matrix at different incubation times in a batch-wise mode. Various mathematic models were employed to explain the amount of adsorbed LPS, and the normalized root mean square error (NRMSE), relative prediction error (RE), and relative percentage error (RPE) were utilized to identify the best-fitting model. Results: The kinetics study revealed that the pseudo-second-order (PSO) reaction, and pore diffusion mass transfer were the rate-controlling steps of LPS adsorption on the S3E3-S-Sepharose and pH of samples did not affect the LPS adsorption kinetics. Conclusion: These findings provide valuable insights for scaling up the LPS removal process through affinity chromatography, contributing to advances in biopharmaceutical and vaccine production research. http://vacres.pasteur.ac.ir/article-1-343-en.pdf 2026-02-17 1 8 10.52547/vacres.10.2.1 Affinity chromatography cationic amphiphilic peptide endotoxin removal kinetic model Mina Sepahi msepahi56@yahoo.com 1 Nano-Biotechnology Department, New Technologies Research Group, Pasteur Institute of Iran, Tehran, 1316943551, Iran. AUTHOR https://orcid.org/0000-0001-9422-8441 Shahin Hadadian hadadian@yahoo.com 2 Nano-Biotechnology Department, New Technologies Research Group, Pasteur Institute of Iran, Tehran, 1316943551, Iran. AUTHOR Reza Ahangari Cohan cohan_r@yahoo.com 3 Nano-Biotechnology Department, New Technologies Research Group, Pasteur Institute of Iran, Tehran, 1316943551, Iran. AUTHOR Dariush Norouzian dariushnorouzain@yahoo.com 4 Nano-Biotechnology Department, New Technologies Research Group, Pasteur Institute of Iran, Tehran, 1316943551, Iran. AUTHOR

SHORT_COMMUNICATION This article explores the subject of mitigating the risks associated with technology transfer in the bioproduction sector such as vaccine industries, which is a challenging and multifaceted process. Based on an examination of case study data, industry reports, and insights gained from over 20 years of bioprocessing experience, it is evident that a comprehensive understanding of the variables involved and meticulous attention to detail are necessary for successful scale-up. Key technological factors include strict regulatory compliance, equipment compatibility, and optimization of process parameters. Organizations can minimize the risks associated with scaling up bioproduction processes and effectively manage the complexities of technology transfer by implementing data-driven strategies and advanced modeling tools. http://vacres.pasteur.ac.ir/article-1-346-en.pdf 2026-02-17 9 10 10.52547/vacres.10.2.9 Commercialization Design Engineering Bioproduction Planning Quality Vahid Marandi vahid.marandi@gmail.com 1 Pasteur Institute of Iran AUTHOR https://orcid.org/0000-0002-9209-7895

OTHERS_CITABLE Introduction: Cutaneous leishmaniasis (CL) due to Leishmania major (L. major) is a widespread vector-borne parasitic infection in subtropical areas. Numerous studies have been conducted to present possible vaccination candidates to address CL. In the present study, 18 L. major vaccine candidate antigens, namely gp46, CatL, CatB, grp78, H1, H2A, H2B, and H4, HSP60, HSP70, HSP83 (HSP90), HSP100, rP0, KMP11, STI-1, TSA, LeIF, and LACK were evaluated by in silico methods to find novel immunogenic epitopes. Methods: online predictions were performed regarding physicochemical, solubility, antigenicity, allergenicity, signal peptide, and transmembrane domains. Since four proteins (i.e., CatB, CatL, gp46, and grp78) were shown to possess signal peptides and transmembrane domains, they were further analyzed along with two antigenic proteins (STI-1 and H2A), regarding post-translational modifications (PTMs), structural (secondary and tertiary) predictions, and epitope mapping for B-cells, cytotoxic T-lymphocyte (CTL) and helper T-lymphocyte (HTL) epitopes against human leukocyte antigen (HLA) reference sets. Results: The world coverage of the CTL and HTL allele-epitope compositions were 96.34% and 41.78%, respectively. Finally, potentially-immunogenic CTL (n = 8) and antigenic HTL (n = 8) epitopes, which were strong IFN-γ inducers, along with 6 B-cell epitopes were selected. Conclusion: These epitopes are potential immunodominant regions among these antigens that upon further evaluations could be considered for a multi-epitope vaccine construction against CL http://vacres.pasteur.ac.ir/article-1-349-en.pdf 2026-02-17 11 27 10.52547/vacres.10.2.11 Leishmania major vaccine candidates Immunoinformatics Immunogenic epitopes Hamed Rezayatmand h.rezayatmand.st@kmu.edu.ua 1 Department of Pharmacy and Technology of Organic Substances, Industrial Pharmacy, Ukrainian state, Chemical Technology University, Dnipro, Ukraine AUTHOR Golnoush Farahzadeh golif29@gmail.com 2 Pharmaceutical chemistry, faculty of chemistry, Azad Tehran north university, Tehran, IRAN AUTHOR Iraj Alipourfard ialipourfard@ichf.edu.pl 3 Institute of physical chemistry, Polish Academy of Science، Warsaw, Poland. AUTHOR Arash Rezaeirad arashrezaeirad@gmail.com 4 Institute of science, Molecular Biology, Üsküdar University, Istanbul, Türkiye AUTHOR Mahsa Alem m.alem@urmia.ac.ir 5 Department of Microbiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran. AUTHOR Sasan Khazaei sasan.kh91@gmail.com 6 Department of Parasitology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran AUTHOR Davood Siamian Davood_Siyamiyan@gmail.com 7 Department of Biology, Faculty of Basic Science, Islamic Azad University, Tonekabon Branch, Mazandaran, Iran AUTHOR Negar Jahani negarjahani@gmail.com 8 Faculty of Veterinary Medicine, Islamic Azad University, Science and Research Branch, Tehran, Iran AUTHOR Faezeh Kalantarzadeh kalantarzadeh98@gmail.com 9 Faculty of Veterinary Medicine, Islamic Azad University, Shushtar Branch, Shushtar, Iran AUTHOR Mohammad Arad Zandieh M.aradzandieh@gmail.com 10 Faculty of Veterinary Medicine, Islamic Azad University, Science and Research Branch, Tehran, Iran AUTHOR Seyyed Amir Hosseini AmirHosseini718@yahoo.com 11 Department of Internal Medicine, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran AUTHOR Saeed Heydarian s.heydarian@gmail.com 12 Department of Internal Medicine, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran AUTHOR Hamidreza Majidiani majidianih1@nums.ac.ir 13 Healthy Aging Research Centre, Neyshabur University of Medical Sciences, Neyshabur, Iran AUTHOR Azar Najafi azar.najafi98@gmail.com 14 Deputy of Research and Technology, Alborz University of Medical Sciences, Karaj, Iran AUTHOR

ORIGINAL_ARTICLE Introduction: Influenza A (H1N1) virus is one of the main causes of seasonal and pandemic influenza in humans. Considering the inherent antigenic changes of the virus, new infectious strains of the virus emerge frequently. In addition, the current methods of vaccine production are mainly based on fully inactivated viruses, which are time-consuming and expensive approaches. In this regard, the development of new protective vaccines will be a reasonable approach. This study aimed to design and express a subunit vaccine based on the hemagglutinin protein of the pH1N1 strain, enriched with protective epitopes of four highly pathogenic virus strains, namely, H1N1, H3N2, H5N1, and H7N9. Methods: Using bioinformatics tools, highly antigenic and conserved epitopes were selected from H1N1, H3N2, H5N1, H7N9 strains and placed at the appropriate positions on HA protein framework of pH1N1 influenza virus. The in silico analyses including immunogenicity, conservancy and population coverage were used to examine the construct, named as Hemag98. The genes sequence was synthesized and transferred to an Escherichia coli host for expression. The expression of Hemag98 protein was then confirmed by SDS-PAGE and Western blotting techniques. The immunogenicity of the purified recombinant proteins was preliminary examined in a mouse model. Results: Based on the predicted results, Hemag98 was shown to be a stable and hydrophilic protein with similar tertiary structure to native HA1 of H1N1 virus. Based on the Z-score of the predicted models, the Hemag98 model was in range of X-ray structured proteins. The expressed protein had the expected 45 KDa molecular weight and the initial in vivo experiment showed that the recombinant Hemag98 protein stimulates specific immune responses. Conclusion: The overall results indicated that the Hemag98 can be considered as a promising platform for the development of a preventive influenza vaccine capable of protecting against diverse strains of influenza virus.   http://vacres.pasteur.ac.ir/article-1-355-en.pdf 2026-02-17 28 38 10.52547/vacres.10.2.28 Influenza In silico Multi-epitope Recombinant vaccine Soheila Hoseini 1 Department of Agricultural Biotechnology, Agricultural College, Tehran University, Karaj Campus, Karaj 31587-77871, Iran AUTHOR Parastoo Ehsani 2 Department of Molecular Biology, Pasteur Institute of Iran, 69 Pasteur Ave., Tehran 1316943551, Iran. 3Department of Influenza and Other Respiratory Viruses, Pasteur Institute of Iran, 69 Pasteur Ave., Tehran 1316943551, Iran AUTHOR Fatemeh Fotuhi 3 Department of Influenza and Other Respiratory Viruses, Pasteur Institute of Iran, 69 Pasteur Ave., Tehran 1316943551, Iran. AUTHOR Behrokh Farahmand 4 Department of Influenza and Other Respiratory Viruses, Pasteur Institute of Iran, 69 Pasteur Ave., Tehran 1316943551, Iran. AUTHOR Houshang Alizadeh 5 Department of Agricultural Biotechnology, Agricultural College, Tehran University, Karaj Campus, Karaj 31587-77871, Iran AUTHOR

OTHERS_CITABLE Introduction: Vaccination against severe acute respiratory syndrome coronavirus (SARS-CoV-2) plays a major role in the global fight against the COVID-19 pandemic. However, the availability of and access to a safe COVID-19 vaccine alone may not be sufficient to curb the epidemic. Vaccine hesitancy—including delays and refusals—remains a major obstacle to achieving herd immunity. This study aimed to investigate public attitudes toward the COVID-19 vaccine. Methods: This descriptive and analytical study was conducted on 1,544 individuals over 30 years of age in South Khorasan Province, selected via convenience sampling through the SIB system. Data were collected using a questionnaire to measure attitudes, administered via telephone interviews. Statistical analyses included mean and standard deviation, frequency and percentage distributions, chi-square tests, independent samples t-tests, and logistic regression analyses, with a significance level set at P < 0.05. Results: The findings revealed variations in vaccination rates across demographic categories, including gender, education level, marital status, occupation, and place of residence. Key factors influencing hesitancy included doubts about the accuracy of vaccine-related information, strong fears of vaccination, and the belief that individuals who had not contracted COVID-19 were naturally resistant and did not require vaccination. Conclusion: Public attitudes, doubts, and concerns regarding the COVID-19 vaccine significantly impact vaccination coverage rates. Therefore, understanding the reasons behind vaccine hesitancy is essential for designing targeted interventions.                                                                                                       http://vacres.pasteur.ac.ir/article-1-348-en.pdf 2026-02-17 39 43 10.52547/vacres.10.2.39 Covid-19 disease Covid-19 vaccine Attitude Zahra Vahdaninya vahdane@bums.ac.ir 1 Social determinants research center, Birjand university of Medical Science , Birjand, Iran AUTHOR https://orcid.org/0000-0003-3119-8200 fatemeh salmani salmany_fatemeh@bums.ac.ir 2 Department of Epidemiology and Biostatics, school of health , social determinants research center, Birjand university of Medical Science , Birjand, Iran AUTHOR https://orcid.org/0000-0001-6781-3899 Fatemeh khosravy fkhosravi4927@bums.ac.ir 3 Social determinants research center, Birjand university of Medical Science , Birjand, Iran AUTHOR https://orcid.org/0000-0002-8655-3938 Fatemeh Baghernezhad hesary ftbagher@gmail.com 4 department of heath education and promotion . school of health , social determinants research center, Birjand university of Medical Science , Birjand, Iran AUTHOR https://orcid.org/0000-0002-2977-4320

SHORT_COMMUNICATION Introduction: Most of the previous researches about Foot and Mouth Disease vaccine have stated that traditional and inactive vaccines cannot comprehensively control the disease and should be reconsidered. Currently, for this reason, the efforts of researchers are on the production and development of new generation vaccines. FMDV VP1 protein is one of the structural proteins and is crucial for the virus’s ability to infect host cells, elicitation of protective immune responses and the determination of serotype specificity. It plays a significant role in the virus’s antigenicity and is a target for vaccine development. Methods: The coding sequence of FMDV VP1 protein (O IR/P50/2016) was extracted from the NCBI site. Recombinant structure designed during heat shock method was transferred into Escherichia coli strain BL21. After culture of transformed bacteria and confirming the presence of the gene by PCR, restriction enzyme digestion and DNA sequencing, the recombinant FMDV VP1 protein was expressed and confirmed by SDS-PAGE and Western blotting. Results: The results of the electrophoresis, PCR and enzymatic digestion indicated that the FMDV VP1 protein gene was correctly inserted and cloned into pET-45 plasmid. SDS-PAGE and Western blotting demonstrated the existence of a protein with a molecular weight of 27 kDa. Conclusion: The results of the current study indicated that the prokaryotically expressed FMDV VP1 protein can potentially be considered as a replacement for old vaccines or antigens (in diagnostic kits) upon further investigations. http://vacres.pasteur.ac.ir/article-1-357-en.pdf 2026-02-17 44 48 10.52547/vacres.10.2.44 Protein Expression E. coli FMD Vaccine candidate Western Blot Atieh Eftekhari aeftekhari2020@ut.ac.ir 1 Department of Microbiology & Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran AUTHOR https://orcid.org/0009-0008-5253-078X Khosrow Aghaeipour Kolyani 2 Department of Biotechnology, Behesht Aein Laboratory Complex, (MABA), Tehran, Iran AUTHOR https://orcid.org/0000-0002-2305-6053 Bahman Abedi Kiasari 3 Department of Microbiology & Immunology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran AUTHOR https://orcid.org/0000-0002-7377-1434 Mohammad Mahdi Ranjbar 4 Department of Foot and Mouth Disease, Razi Vaccine and Serum Research Institute, Karaj, Iran AUTHOR Homayoun Mahravani 5 Department of Foot and Mouth Disease, Razi Vaccine and Serum Research Institute, Karaj, Iran AUTHOR https://orcid.org/0000-0002-9802-2034

OTHERS_CITABLE Introduction: Influenza, particularly Influenza A, remains a significant global health challenge due to its high morbidity and mortality rates. The hemagglutinin (HA) protein plays a critical role in viral entry, replication, and disease progression, while neuraminidase (NA) is essential for the forming and releasing of viral particles. Despite the ongoing challenge of HA protein variability, which leads to the emergence of new antigenic strains, this study focuses on enhancing antibody responses and achieving partial protection against influenza through the use of HA and NA proteins. Methods: A recombinant peptide vaccine targeting Influenza A was developed and tested in mice, showing strong immune responses and increased protection. Evaluated through computational, laboratory, and animal studies, the vaccine proved effective both alone and with an adjuvant. Prior bioinformatics analyses ensured its safety and stability, highlighting the promise of using immunodominant epitopes in recombinant vaccines for better influenza prevention. Results: The recombinant peptide vaccine against Influenza A viruses demonstrated strong immune responses and enhanced protection in a mice model. Evaluations under in silico, in vitro, and in vivo conditions confirmed the vaccine’s efficacy, both alone and with an adjuvant, through robust immune responses and cytokine production. These findings highlight the potential of bioinformatically guided immunodominant epitopes in recombinant vaccines for improved influenza control. Conclusion: This recombinant vaccine prototype shows strong immunogenicity and potential for effective influenza control in mice model. http://vacres.pasteur.ac.ir/article-1-358-en.pdf 2026-02-17 49 55 10.52547/vacres.10.2.49 Influenza virus Multi-epitope Peptide vaccine Immunogenicity Mina Mirzaee mirzaie.mina@ymail.com 1 Department of Microbiology and Microbial Biotechnology, Faculty of Life Science and Biotechnology, University of Shahid Beheshti, Tehran, Iran AUTHOR Fatemeh Fotouhi 2 Department of Influenza and Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran AUTHOR Hadiseh Shokohi 3 Department of Influenza and Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran AUTHOR Parisa Roshani Asl 4 Department of Influenza and Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran AUTHOR Hadi Bamehr 5 Department of Influenza and Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran AUTHOR Golnaz Bahramali 6 Department of Hepatitis and AIDS, Pasteur Institute of Iran, Tehran, Iran. AUTHOR Behrokh Farhmand 7 Department of Influenza and Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran AUTHOR https://orcid.org/0000-0002-7772-3595 Seyed Masoud Hosseini 8 Department of Microbiology and Microbial Biotechnology, Faculty of Life Science and Biotechnology, University of Shahid Beheshti, Tehran, Iran AUTHOR

REVIEW_ARTICLE Conventional lung cancer therapies—including surgery, chemotherapy, and radiotherapy—are often limited by insufficient efficacy and significant adverse effects. In response, cancer immunotherapy, particularly vaccine-based approaches, has emerged as a promising strategy to improve outcomes. Carbon nanotubes (CNTs) represent a versatile nanoplatform capable of efficiently delivering antigens, adjuvants, and therapeutic agents, making them attractive candidates for nanovaccine development. This review examines the role of CNT-based nanovaccines in lung cancer immunotherapy, focusing on their mechanisms of action and therapeutic potential. Evidence indicates that CNT nanovaccines enhance antitumor immunity by promoting dendritic cell maturation, stimulating robust humoral and cellular immune responses, and reprogramming the immunosuppressive tumor microenvironment toward an immunopermissive state. Despite these promising preclinical results, clinical translation requires further optimization of CNT design and functionalization, alongside comprehensive in vivo evaluation of immunogenicity, biodistribution, and safety profiles. Overall, CNT-based nanovaccines hold considerable potential to advance lung cancer immunotherapy, though targeted research is needed to bridge the gap between experimental promise and clinical application. http://vacres.pasteur.ac.ir/article-1-366-en.pdf 2026-02-17 56 59 10.52547/vacres.10.2.56 Lung cancer carbon nanotube nanovaccine immunotherapy Mojgan Sheikhpour m_sheikhpour@pasteur.ac.ir 1 Department of Mycobacteriology and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran. 2Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran AUTHOR https://orcid.org/0000-0002-1927-6555 Hanie Sakhi 2 Department of Mycobacteriology and Pulmonary Research, Pasteur Institute of Iran, Tehran, Iran. 2Microbiology Research Center (MRC), Pasteur Institute of Iran, Tehran, Iran AUTHOR