{
  "abstract": "The rapid growth of recombinant protein vaccines underscores the need for manufacturing platforms that are efficient, stable, and scalable. We present a biomanufacturing approach integrating bioengineered Pichia pastoris with sodium alginate bioinks using the patented Sprayed Multi Adsorbed-Droplet Reposing Technology (SMART, US20250288524A1) to fabricate uniform microbiofactories. Engineered Pichia pastoris X33 strains expressing the truncated Epstein–Barr virus nuclear antigen 1 (EBNA1, aa 379–641) from the pPICZαA-EBNA1 vector were encapsulated and induced with methanol for antigen production. SMART-assisted encapsulation generated microcarriers averaging ∼50 μm in diameter with a postencapsulation yeast recovery of 87% as assessed by the Alamar Blue viability assay. Across three independent biological replicates, hydrogel-encapsulated yeast consistently produced higher EBNA1 titers than free yeast, corresponding to a 1.70 ± 0.37-fold enhancement, as quantified by ELISA and confirmed by Western blot. Following lyophilization, in a single-experiment proof-of-concept storage study, the microbiofactories sustained EBNA1 production at 392 ± 11 ng/mL over 28 days at ambient temperature (24 °C), compared to 154 ± 18 ng/mL for free yeast; the sustained ELISA signal at day 28 constitutes functional evidence of metabolically active yeast upon reconstitution. These findings demonstrate the in vitro proof-of-concept feasibility of SMART-assisted hydrogel encapsulation as an ambient-temperature-stable platform for microbial antigen production. This approach provides a foundation for future development of biomanufacturing systems that may help reduce cold-chain dependence, pending further scale-up and immunological validation.",
  "abstract_shown": true,
  "affiliations": [
    "University of Mississippi , , , ,"
  ],
  "article_type": "research article",
  "arxiv": null,
  "author_total": 5,
  "authors": [
    {
      "affs": [
        0
      ],
      "name": "Shao-Ching Feng",
      "orcid": "0009-0001-3898-4880"
    },
    {
      "affs": [
        0
      ],
      "name": "Prashant Pandey"
    },
    {
      "affs": [
        0
      ],
      "name": "Leela Raghava Jaidev Chakka",
      "orcid": "0000-0003-0039-0064"
    },
    {
      "affs": [
        0
      ],
      "name": "Yu Zhang",
      "orcid": "0000-0002-8925-2284"
    },
    {
      "affs": [
        0
      ],
      "name": "Mohammed Maniruzzaman",
      "orcid": "0000-0002-3373-5586"
    }
  ],
  "bibtex": "https://research.draconitic.com/paper/8hz1teqqfr1j7.bib",
  "cited_by": 0,
  "doi": "10.1021/acsomega.6c05614",
  "fulltext_url": "https://doi.org/10.1021/acsomega.6c05614",
  "id": "8hz1teqqfr1j7",
  "issns": [
    "2470-1343"
  ],
  "landing_url": "https://pubs.acs.org/acsodf/article/doi/10.1021/acsomega.6c05614/5454859/SMART-Assisted-Fabrication-of-Engineered-Pichia",
  "language": "en",
  "licence": "CC-BY-NC-ND-4.0",
  "licence_label": "CC BY-NC-ND 4.0",
  "licence_source": "article",
  "licence_url": "https://creativecommons.org/licenses/by-nc-nd/4.0/",
  "metadata_licence": null,
  "native_subjects": [
    [
      "heading",
      "Article"
    ],
    [
      "lcc",
      "QD1-999"
    ],
    [
      "openalex",
      "1312|Fungal and yeast genetics research"
    ]
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  "openalex": "W7223153500",
  "pdf_url": "https://pubs.acs.org/doi/pdf/10.1021/acsomega.6c05614",
  "pmcid": "PMC13737990",
  "pmid": "42992052",
  "published": "2026-10-01",
  "publisher": "American Chemical Society (ACS)",
  "ris": "https://research.draconitic.com/paper/8hz1teqqfr1j7.ris",
  "sources": [
    {
      "first_seen": "2026-10-30T12:18:39Z",
      "native_id": "10.1021/acsomega.6c05614",
      "source": "crossref",
      "url": "https://pubs.acs.org/acsodf/article/doi/10.1021/acsomega.6c05614/5454859/SMART-Assisted-Fabrication-of-Engineered-Pichia"
    },
    {
      "first_seen": "2026-11-04T14:20:38Z",
      "native_id": "12f3e043987748ab855ec5bbfbc7fb99",
      "source": "doaj",
      "url": "https://doi.org/10.1021/acsomega.6c05614"
    },
    {
      "first_seen": "2026-11-09T12:23:04Z",
      "native_id": "PMC13737990",
      "source": "pmc",
      "url": "https://doi.org/10.1021/acsomega.6c05614"
    }
  ],
  "subjects": [
    {
      "field": "biochemistry",
      "origin": "openalex:1312|Fungal and yeast genetics research",
      "subfield": "molecular-biology"
    }
  ],
  "title": "SMART-Assisted Fabrication of Engineered Pichia pastoris Microbiofactories for Stable In Vitro Antigen Production and Storage",
  "title_text": "SMART-Assisted Fabrication of Engineered Pichia pastoris Microbiofactories for Stable In Vitro Antigen Production and Storage",
  "type": "peer-reviewed",
  "updated_at": "2026-11-09T12:23:04Z",
  "url": "https://research.draconitic.com/paper/8hz1teqqfr1j7",
  "venue": "ACS Omega",
  "versions": []
}