Browsing by Subject "Antibody"

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    Ant i-neuronal and stress-induced-phosphoprotein 1 antibodies in neuro-Behcet's disease
    (Elsevier, 2011-10-28) Vural, B.; Uğurel, E.; Tüzün, E.; Kürtüncü, M.; Zuliani, L.; Çavus, F.; İçoz, S.; Erdağ, E.; Gül, A.; Güre, A. O.; Vincent, A.; Özbek, U.; Eraksoy, M.; Demir, G. A.
    No disease-specific neuronal antibodies have so far been defined in neuro-Behçet's disease (NBD). Immunohistochemistry and immunocytochemistry studies showed antibodies to hippocampal and cerebellar molecular layers and the surface antigens of cultured hippocampal neurons in sera and/or cerebrospinal fluids (CSF) of 13 of 20 NBD and 6 of 20 BD patients but not in multiple sclerosis or headache controls. Screening with a protein macroarray led to identification of stress-induced-phosphoprotein-1 (STIP-1) as an antigenic target. High-titer STIP-1-antibodies were detected in 6 NBD patients' sera but not in controls. These results suggest that neuronal antibodies could be useful as diagnostic biomarkers in NBD. © 2011 Elsevier B.V.
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    Large-scale manufacturing and characterization of a Sars Cov-2 virus-like particle vaccine adsorbed onto alhydrogel and adjuvanted with K3 CpG oligodeoxynucleotide for use in phase 1/2 clinical trials
    (2022-04-28) Bülbül, Artun
    Emergence of COVID-19 pandemic has been met by an exceptionally fast response from vaccine makers around the globe. Vaccines that elicit excellent immunological responses against SARS-CoV-2 are now widely utilized. Existing platforms include mRNA-lipid nanoparticle-based vaccines, adenovirus vectored vaccines, various inactivated virus vaccines and subunit vaccines. We have previously described a novel virus-like particle (VLP) platform expressing the hexaproline prefusion stabilized Spike protein along with the nucleocapsid, membrane and envelope structural proteins. In mice, ferrets and rats, VLPs adjuvanted with K3 CpG Oligodeoxynucleotide (ODN) and adsorbed onto 2% Aluminum Hydroxide (Alum), induced robust humoral and cellular immune response against Spike and Nucleocapsid proteins. Herein, we have expanded our work to manufacture the virus like particles in a GMP compliant facility intended for testing in phase I/II clinical trials. The technology transfer comprises i) VLP production from suspension adapted HEK293 cells, ii) purification with multimodal fast protein liquid chromatography (FPLC) and iii) concentration and diafiltration using tangential flow filtration (TFF). We have successfully scaled up our production from 50 mL of HEK293 cell culture to 5 L bioreactor, achieving yields reaching up to 40 mg VLPs per L of cell culture. Furthermore, several methods were developed to determine protein identity, purity, functionality, stability and immunopotency of VLP vaccine that was finally formulated with Alum + CpG ODN. Moreover, we investigated the immunogenicity of VLPs decorated either with Wuhan (Hu-1) or with Alpha (B.1.1.7) variant Spike against receptor binding domains (RBD) specific to other variants of concern (VoC). Although our vaccine platform, could further benefit from process optimization to improve VLP yield, this study presents the first pilot scale production and purification of variant specific hexaproline prefusion stabilized SARS-CoV-2 VLPs. VLP preparations complying with our quality control parameters were released for fill and finish and were used for subsequent Phase 1 (NCT04818281) and Phase 2 clinical trials (NCT04962893).
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    Mitochondrial carrier homolog 1 (Mtch1) antibodies in neuro-Behçet's disease
    (Elsevier, 2013) Vural, B.; Şehitoğlu, E.; Çavuş, F.; Yalçınkaya, N.; Haytural, H.; Küçükerden, M.; Ulusoy, C.; Uğurel, E.; Turan, S.; Bulut, L.; Türkoğlu, R.; Shugaiv, E.; Kürtüncü, M.; Atakan, S.; Güre, A. O.; Gül, A.; Eraksoy, M.; Demir, G. A.; Tüzün, E.
    Efforts for the identification of diagnostic autoantibodies for neuro-Behcet's disease (NBD) have failed. Screening of NBD patients' sera with protein macroarray identified mitochondrial carrier homolog 1 (Mtch1), an apoptosis-related protein, as a potential autoantigen. ELISA studies showed serum Mtch1 antibodies in 68 of 144 BD patients with or without neurological involvement and in 4 of 168 controls corresponding to a sensitivity of 47.2% and specificity of 97.6%. Mtch1 antibody positive NBD patients had more attacks, increased disability and lower serum nucleosome levels. Mtch1 antibody might be involved in pathogenic mechanisms of NBD rather than being a coincidental byproduct of autoinflammation. © 2013 Elsevier B.V.
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    Performance comparison of aptamer- and antibody-based biosensors for bacteria detection on glass surfaces
    (Taylor & Francis Inc., 2025) Balcı, O.; Kürekçi, A.; Özalp, V. C.; Barbaros, Çetin
    Antibodies are the most common ligands in commercial and research assay systems for detecting whole pathogen cells. On the other hand, aptamers are superior ligands with many advantages over antibodies in sensitive and robust assay development. Extensive comparisons between aptamer-based biosensors and immune sensors are limited to protein analytes. Here, we report a comparison of ligands (four antibodies and one aptamer for each bacteria) to be used as a biosensor for Escherichia coli and Staphylococcus aureus on glass surfaces through systematic experiments. We have demon-strated that anti-E. coli antibody and mouse monoclonal to S. aureus have the best performance among the compared ligands. Hence, the ligands with the best performance were further investigated within the scope of linear range, analytical sensitivity, and reproducibility of the results. We have demonstrated that anti-E. coli antibody with a capture efficiency of 89.1% and mouse monoclonal to S. aureus with a capture efficiency of 88.2% have the best performance among the compared ligands. The results suggest that antibody ligands function with higher efficiency than aptamer ligands but aptamers have strong potential as an analytical tool.
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    Protocol to study neuronal membrane proteasome function in mouse peripheral sensory neurons
    (Cell Press, 2025-03-21) Krueger, Emily R.; Church, Taylor R.; Brennan, Anna; Türker, Fulya; Villalón Landeros, Eric
    Neuronal membrane proteasomes (NMPs) are expressed on a subset of somatosensory dorsal root ganglion (DRG) neurons and influence mechanical and pain sensitivity. Here, we present a protocol for studying NMP function in mouse peripheral sensory neurons. We describe steps for procuring and culturing primary DRG neurons. We then detail biochemical and antibody feeding approaches to analyze NMP expression and localization. Finally, we include Ca2+ imaging techniques for investigating NMP function in DRG neurons. For complete details on the use and execution of this protocol, please refer to Villalón Landeros et al.1 © 2024 The Author(s)