Vaccine Building ‘Kit’: Combining Peptide Bricks to Elicit A Desired Immune Response Without Adding An Adjuvant

Gorbunov, Aleksandr A; Sannikova, E. P.; Gubaidullin, I. I.; Serobyan, Gayane A; Gorbunova, A. Yu.; Serkina, A. V.; Plokhikh, Konstantin S.; Kamyshinsky, Roman; Vorovitch, Mikhail F.; Bulushova, N. V.; Kuchin, Sergei; Козлов, Д. Г.

doi:10.2217/nnm-2021-0424

Cited by 1 publication

(8 citation statements)

References 43 publications

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“…Some plasmids used in this work were constructed earlier (Gorbunov et al, 2022), including the following:…”

Section: Methodsmentioning

confidence: 99%

“…Transformants were grown at 37°C in LB medium supplemented with kanamycin (30 μg/mL) on a rotary shaker at 250 rpm. Some plasmids used in this work were constructed earlier (Gorbunov et al, 2022), including the following: Plasmid pET28s is a derivative of the pET28b+ (Novagen) containing a unique SalI site replacing the unique BglII site in pET28b+.…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…A unique BamHI site separates the sequences encoding SUMO and His 6 in these plasmids. Plasmid pET28s-L 6 KD-SUMO-PhtD19 was used for the synthesis of the L 6 KD-SUMO-PhtD19 protein; this protein contains the PhtD19 peptide from pneumococcal histidine triad protein D (a.a. 200-219) (Papastamatiou et al, 2018) that is of interest as a vaccine component (Gorbunov et al, 2022).…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…Here, we describe a new platform for synthesizing target proteins/peptides in the form of AIBs. It inherits the design of L 6 KD‐SUMO fusions used for highly immunogenic nanoparticle generation presented in our previous work (Gorbunov et al, 2022). In the current study, a wide range of proteins and peptides were produced in E. coli cells using the L 6 KD‐SUMO platform, and the attention was focused on investigation of the platform‐directed protein structures expressed in vivo “as is,” without renaturation.…”

Section: Introductionmentioning

confidence: 98%

“…Here, we describe a new platform for synthesizing target proteins/peptides in the form of AIBs. It inherits the design of L 6 KD-SUMO fusions used for highly immunogenic nanoparticle generation presented in our previous work (Gorbunov et al, 2022).…”

mentioning

confidence: 99%

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Synthesis of biologically active proteins as L6KD‐SUMO fusions forming inclusion bodies in Escherichia coli

Komolov,

Sannikova,

Gorbunov

et al. 2023

Biotech & Bioengineering

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A new platform has been developed to facilitate the production of biologically active proteins and peptides in Escherichia coli. The platform includes an N‐terminal self‐associating L6KD peptide fused to the SUMO protein (small ubiquitin‐like protein modifier) from the yeast Saccharomyces cerevisiae, which is known for its chaperone activity. The target proteins are fused at the C termini of the L6KD‐SUMO fusions, and the resulting three‐component fusion proteins are synthesized and self‐assembled in E. coli into so‐called active inclusion bodies (AIBs). In vivo, the L6KD‐SUMO platform facilitates the correct folding of the target proteins and directs them into AIBs, greatly simplifying their purification. In vitro, the platform facilitates the effective separation of AIBs by centrifugation and subsequent target protein release using SUMO‐specific protease. The properties of the AIBs were determined using five proteins with different sizes, folding efficiencies, quaternary structure, and disulfide modifications. Electron microscopy shows that AIBs are synthesized in the form of complex fibrillar structures resembling “loofah sponges” with unusually thick filaments. The obtained results indicate that the new platform has promising features and could be developed to facilitate the synthesis and purification of target proteins and protein complexes without the use of renaturation.

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“…Some plasmids used in this work were constructed earlier (Gorbunov et al, 2022), including the following:…”

Section: Methodsmentioning

confidence: 99%

Section: Plasmid Constructionmentioning

confidence: 99%