Science & Insights

@article{Diaz2025,

title = {SARS-CoV-2 spike peptide analysis reveals a highly conserved region that elicits potentially pathogenic autoantibodies: implications to pan-coronavirus vaccine development},

author = {Marilyn Diaz and Zbignew Mikulski and Dan Leaman and Angel Gandarilla and Nathalia Da Silva and Annie Verkoczy and Jinsong Zhang and Laurent Verkoczy},

doi = {10.3389/fimmu.2025.1488388},

issn = {1664-3224},

year  = {2025},

date = {2025-02-25},

urldate = {2025-02-25},

journal = {Front. Immunol.},

volume = {16},

publisher = {Frontiers Media SA},

abstract = {The SARS-CoV-2 pandemic, while subsiding, continues to plague the world as new variants emerge. Millions have died, and millions more battle with the debilitating symptoms of a clinical entity known as long Covid. The biggest challenge remains combating an ever-changing variant landscape that threatens immune evasion from vaccine and prior infection-generated immunity. In addition, the sequelae of symptoms associated with long Covid almost certainly point to multiple pathologies that range from direct damage to organs during infection to a potential role for infection-induced autoreactive antibodies in promoting autoimmune-like conditions in these patients. In this study, a peptide scan of the SARS-CoV-2 spike protein was done to detect novel, highly conserved linear epitopes that do not elicit autoantibodies. We identified eight predicted linear epitopes capable of eliciting anti-spike IgG antibodies. Immunizations alternating peptide conjugated to KLH with the full trimer yielded the highest antibody levels, but homologous immunization with some of the peptides also yielded high levels when an additional immunization step was added. Of all regions tested, the stem helix adjacent to the heptad repeat 2 (HR2) region also elicited high levels of autoreactive antibodies to known autoantigens in common systemic autoimmune disorders such as lupus and scleroderma and may contribute to the long Covid syndrome seen in some patients. Implications to vaccine design are discussed.},

keywords = {coronavirus, SARS-CoV-2},

pubstate = {published},

tppubtype = {article}

}

@article{Mosca2025,

title = {Standardization of hemoglobin A2 and hemoglobin F: Achievements and perspectives},

author = {Andrea Mosca and Cristian Arsene and Renata Paleari and Patricia Kaiser and Kees Harteveld and Yvonne Daniel and Chie Amano and Atsushi Murakami and Guy Auclair},

doi = {10.1016/j.cca.2024.120087},

issn = {0009-8981},

year  = {2025},

date = {2025-02-00},

urldate = {2025-02-00},

journal = {Clinica Chimica Acta},

volume = {567},

publisher = {Elsevier BV},

abstract = {The establishment of reference systems for the standardization of hemoglobin A2 (HbA2) and fetal hemoglobin (HbF), both critical for improving diagnostic accuracy in conditions such as β-thalassemia and sickle cell disease, are described. Efforts were led by the IFCC and other groups to address and reduce the variability in laboratory measurements of these hemoglobins. This document outlines the production of certified reference materials (CRMs) for HbA2 and the development of a reference measurement procedure using isotope dilution mass spectrometry. Similarly, standardizing HbF is essential for supporting diagnostic and therapeutic strategies, particularly in managing sickle cell disease. HbF levels can predict disease outcomes and guide treatment plans. Significant challenges remain in achieving consistent measurement across laboratories, and the process for standardization for this minor hemoglobin has just begun. We are confident that the implementation of these reference systems will provide improved accuracy and traceability in the future.},

keywords = {HbA2, HbF, hemoglobin},

pubstate = {published},

tppubtype = {article}

}

@article{Reis-Claro2024,

title = {Application of the iPLUS non-coding sequence in improving biopharmaceuticals production},

author = {Inês Reis-Claro and Maria Inês Silva and Ana Moutinho and Beatriz C. Garcia and Isabel Pereira-Castro and Alexandra Moreira},

url = {https://trenzyme.com/protein-production-services/custom-protein-expression-service/, Custom Protein Expression Service},

doi = {10.3389/fbioe.2024.1355957},

issn = {2296-4185},

year  = {2024},

date = {2024-02-06},

urldate = {2024-02-06},

journal = {Front. Bioeng. Biotechnol.},

volume = {12},

publisher = {Frontiers Media SA},

abstract = {The biotechnological landscape has witnessed significant growth in biological therapeutics particularly in the field of recombinant protein production. Here we investigate the function of 3′UTR cis-regulatory elements in increasing mRNA and protein levels in different biological therapeutics and model systems, spanning from monoclonal antibodies to mRNA vaccines. We explore the regulatory function of iPLUS - a universal sequence capable of consistently augmenting recombinant protein levels. By incorporating iPLUS in a vector to express a monoclonal antibody used in immunotherapy, in a mammalian cell line used by the industry (ExpiCHO), trastuzumab production increases by 2-fold. As yeast Pichia pastoris is widely used in the manufacture of industrial enzymes and pharmaceuticals, we then used iPLUS in tandem (3x) and iPLUSv2 (a variant of iPLUS) to provide proof-of-concept data that it increases the production of a reporter protein more than 100-fold. As iPLUS functions by also increasing mRNA levels, we hypothesize that these sequences could be used as an asset in the mRNA vaccine industry. In fact, by including iPLUSv2 downstream of Spike we were able to double its production. Moreover, the same effect was observed when we introduced iPLUSv2 downstream of MAGEC2, a tumor-specific antigen tested for cancer mRNA vaccines. Taken together, our study provides data (TLR4) showing that iPLUS may be used as a valuable asset in a variety of systems used by the biotech and biopharmaceutical industry. Our results underscore the critical role of non-coding sequences in controlling gene expression, offering a promising avenue to accelerate, enhance, and cost-effectively optimize biopharmaceutical production processes.},

keywords = {iPLUS, pTZ-p04_GFP},

pubstate = {published},

tppubtype = {article}

}

@article{Horvath2023,

title = {Novel intranasal vaccine targeting SARS-CoV-2 receptor binding domain to mucosal microfold cells and adjuvanted with TLR3 agonist Riboxxim™ elicits strong antibody and T-cell responses in mice},

author = {Dennis Horvath and Nigel Temperton and Martin Mayora-Neto and Kelly Da Costa and Diego Cantoni and Reinhold Horlacher and Armin Günther and Alexander Brosig and Jenny Morath and Barbara Jakobs and Marcus Groettrup and Heinz Hoschuetzky and Jacques Rohayem and Jan ter Meulen},

url = {https://shop.trenzyme.com/products/spike-s1-rbd-liquid, ➥Mentioned protein in trenzyme webshop (SKU P2020-001)},

doi = {10.1038/s41598-023-31198-3},

issn = {2045-2322},

year  = {2023},

date = {2023-12-00},

urldate = {2023-12-00},

journal = {Sci Rep},

volume = {13},

number = {1},

publisher = {Springer Science and Business Media LLC},

abstract = {SARS-CoV-2 continues to circulate in the human population necessitating regular booster immunization for its long-term control. Ideally, vaccines should ideally not only protect against symptomatic disease, but also prevent transmission via asymptomatic shedding and cover existing and future variants of the virus. This may ultimately only be possible through induction of potent and long-lasting immune responses in the nasopharyngeal tract, the initial entry site of SARS-CoV-2. To this end, we have designed a vaccine based on recombinantly expressed receptor binding domain (RBD) of SARS-CoV-2, fused to the C-terminus of C. perfringens enterotoxin, which is known to target Claudin-4, a matrix molecule highly expressed on mucosal microfold (M) cells of the nasal and bronchial-associated lymphoid tissues. To further enhance immune responses, the vaccine was adjuvanted with a novel toll-like receptor 3/RIG-I agonist (Riboxxim™), consisting of synthetic short double stranded RNA. Intranasal prime-boost immunization of mice induced robust mucosal and systemic anti-SARS-CoV-2 neutralizing antibody responses against SARS-CoV-2 strains Wuhan-Hu-1, and several variants (B.1.351/beta, B.1.1.7/alpha, B.1.617.2/delta), as well as systemic T-cell responses. A combination vaccine with M-cell targeted recombinant HA1 from an H1N1 G4 influenza strain also induced mucosal and systemic antibodies against influenza. Taken together, the data show that development of an intranasal SARS-CoV-2 vaccine based on recombinant RBD adjuvanted with a TLR3 agonist is feasible, also as a combination vaccine against influenza.},

keywords = {P2020-001, SARS-CoV-2},

pubstate = {published},

tppubtype = {article}

}

@article{Kovacs2023,

title = {Veklury® (remdesivir) formulations inhibit initial membrane‐coupled events of SARS‐CoV‐2 infection due to their sulfobutylether‐β‐cyclodextrin content},

author = {Tamas Kovacs and Kitti Kurtan and Zoltan Varga and Peter Nagy and Gyorgy Panyi and Florina Zakany},

url = {https://trenzyme.shop/products/spike-s1-rbd-gfp-his-tag, ➥Mentioned protein in trenzyme webshop (SKU P2020-031)

https://trenzyme.shop/products/s1-rbd-delta-b16172-india-gfp-his-tag, ➥Mentioned protein in trenzyme webshop (SKU P2020-049)

https://trenzyme.shop/products/s1-rbd-omicron-b11529-ba1-gfp-his-tag, ➥Mentioned protein in trenzyme webshop (SKU P2020-061)},

doi = {10.1111/bph.16063},

issn = {1476-5381},

year  = {2023},

date = {2023-08-00},

urldate = {2023-08-00},

journal = {British J Pharmacology},

volume = {180},

number = {16},

pages = {2064--2084},

publisher = {Wiley},

abstract = {Background and Purpose

Despite its contradictory clinical performance, remdesivir (Veklury®) has a pivotal role in COVID-19 therapy. Possible contributions of the vehicle, sulfobutylether-β-cyclodextrin (SBECD) to Veklury® effects have been overlooked. The powder and solution formulations of Veklury® are treated equivalently despite their different vehicle content. Our objective was to study Veklury® effects on initial membrane-coupled events of SARS-CoV-2 infection focusing on the cholesterol depletion-mediated role of SBECD.



Experimental Approach

Using time-correlated flow cytometry and quantitative three-dimensional confocal microscopy, we studied early molecular events of SARS-CoV-2–host cell membrane interactions.



Key Results

Veklury® and different cholesterol-depleting cyclodextrins (CDs) reduced binding of the spike receptor-binding domain (RBD) to ACE2 and spike trimer internalization for Wuhan-Hu-1, Delta and Omicron variants. Correlations of these effects with cholesterol-dependent changes in membrane structure and decreased lipid raft-dependent ACE2–TMPRSS2 interaction establish that SBECD is not simply a vehicle but also an effector along with remdesivir due to its cholesterol-depleting potential. Veklury® solution inhibited RBD binding more efficiently due to its twice higher SBECD content. The CD-induced inhibitory effects were more prominent at lower RBD concentrations and in cells with lower endogenous ACE2 expression, indicating that the supportive CD actions can be even more pronounced during in vivo infection when viral load and ACE expression are typically low.



Conclusion and Implications

Our findings call for the differentiation of Veklury® formulations in meta-analyses of clinical trials, potentially revealing neglected benefits of the solution formulation, and also raise the possibility of adjuvant cyclodextrin (CD) therapy, even at higher doses, in COVID-19.},

keywords = {P2020-031, P2020-049, P2020-061, SARS-CoV-2},

pubstate = {published},

tppubtype = {article}

}