1 Ocak 2024 Dergi makalesi Açık Erişim

Gul, Aytul; Alak, Sedef Erkunt; Can, Huseyin; Karakavuk, Muhammet; Korukluoglu, Gulay; Altas, Ayse Basak; Gul, Ceren; Karakavuk, Tugba; Koseoglu, Ahmet Efe; Polat, Hivda Ulbegi; Malkocoglu, Hilal Yazici; Ekiz, Arzu Tas; Abaci, Irem; Aksoy, Ozge; Enul, Hakan; Adiay, Cumhur; Uzar, Serdar; Sarac, Fahriye; Un, Cemal; Guruz, Adnan Yuksel

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    <subfield code="a">&lt;p&gt;Severe acute respiratory syndrome coronavirus 2 had devastating consequences for human health. Despite the introduction of several vaccines, COVID-19 continues to pose a serious health risk due to emerging variants of concern. DNA vaccines gained importance during the pandemic due to their advantages such as induction of both arms of immune response, rapid development, stability, and safety profiles. Here, we report the immunogenicity and protective efficacy of a DNA vaccine encoding spike protein with D614G mutation (named pcoSpikeD614G) and define a large-scale production process. According to the in vitro studies, pcoSpikeD614G expressed abundant spike protein in HEK293T cells. After the administration of pcoSpikeD614G to BALB/c mice through intramuscular (IM) route and intradermal route using an electroporation device (ID + EP), it induced high level of anti-S1 IgG and neutralizing antibodies (P &amp;lt; 0.0001), strong Th1-biased immune response as shown by IgG2a polarization (P &amp;lt; 0.01), increase in IFN-gamma levels (P &amp;lt; 0.01), and increment in the ratio of IFN-gamma secreting CD4+ (3.78-10.19%) and CD8+ (5.24-12.51%) T cells. Challenging K18-hACE2 transgenic mice showed that pcoSpikeD614G administered through IM and ID + EP routes conferred 90-100% protection and there was no sign of pneumonia. Subsequently, pcoSpikeD614G was evaluated as a promising DNA vaccine candidate and scale-up studies were performed. Accordingly, a large-scale production process was described, including a 36 h fermentation process of E. coli DH5 alpha cells containing pcoSpikeD614G resulting in a wet cell weight of 242 g/L and a three-step chromatography for purification of the pcoSpikeD614G DNA vaccine.&lt;/p&gt;</subfield>
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