BBa_257B84Y6: Chain A, Ferritin [Helicobacter pylori J99]

This part encodes ferritin, an engineered protein scaffold designed to self-assemble into uniform, spherical nanoparticles (12–16 nm in diameter). In our project, ferritin serves as a multivalent display platform for conserved influenza hemagglutinin (HA) epitopes, dramatically enhancing their immunogenicity and stability.

When fused to target antigens, the ferritin scaffold enables repetitive, high-density antigen display, mimicking viral architectures to boost immune responses. Our characterization confirmed successful nanoparticle formation and robust structural integrity by both nano flow cytometry and TEM.

Importantly, ferritin-based nanoparticles protect fused antigens from proteolytic degradation and environmental stress, improving storage stability and making them highly suitable for vaccine development, especially in settings lacking cold-chain infrastructure.

We characterized the resulting nanoparticles using two main techniques: nano flow cytometry (NanoFCM) and transmission electron microscopy (TEM). NanoFCM measurements revealed a consistent particle size distribution, while TEM imaging confirmed that the nanoparticles were uniform and spherical, with diameters ranging from 12 to 16 nm—matching our design expectations.

Fig 12. Nano Flow Cytometry Measurement of Ferritin Particles

Fig 13. Transmission Electron Microscopy (TEM) Analysis of Recombinant Ferritin Nanoparticles

Scale: 100mm
Representative TEM images showing the morphology of recombinant ferritin-based nanoparticles. The particles exhibit uniform, spherical structures with diameters ranging from 12 to 16 nm, consistent with the expected theoretical size for ferritin nanoparticles.

BBa_257RV4NA: B-(16)₄ Conserved Influenza B HA Epitope-Ferritin Fusion

B-(16)₄ is a recombinant fusion protein part designed for universal influenza vaccine development. It consists of a highly conserved 16-amino-acid epitope from the HA2 region of influenza B hemagglutinin (HA), fused to a ferritin nanoparticle scaffold. The selected epitope sequence (TISSQIELAVLLSNEC) was computationally identified for strong conservation and antigenic potential, ensuring broad reactivity across influenza B virus strains.

BBa_257RV4NA: H1-(16)₄ – Conserved H1N1 HA Epitope Module

Contains four tandem repeats of a highly conserved 16-amino-acid sequence (DIWTYNAELLVLLENE) derived from the HA2 domain of influenza A H1N1 hemagglutinin. This sequence was selected based on comprehensive bioinformatic analysis for its exceptional conservation and predicted strong antigenicity, ensuring broad coverage across H1N1 strains.

BBa_25KXFCRG: H3-H1-B – Multi-Epitope Conserved Influenza HA Antigen Module

H3-H1-B is a synthetic multi-epitope antigen module engineered for broad-spectrum influenza vaccine development and nanoparticle-based immunogen strategies.

This part contains three tandemly linked, highly conserved 16-amino-acid sequences, each from a different influenza virus subtype:

• H3: Conserved HA2 epitope from influenza A H3 subtype
• H1: Conserved HA2 epitope from influenza A H1 subtype (DIWTYNAELLVLLENE)
• B: Conserved HA2 epitope from influenza B subtype (TISSQIELAVLLSNEC)