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Construction, expression and functional characterization of an engineered antibody against tumor antigen MUC-1C

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  • Mona Pourjafar, Research Center for Molecular, Aarhus Universitet
  • ,
  • Michaela Miehe
  • Rezvan Najafi, Research Center for Molecular
  • ,
  • Meysam Soleimani, School of Pharmacy
  • ,
  • Edzard Spillner

Minibodies (single-chain Fv-CH3) are fusion proteins of a single-chain variable fragment (scFv) to the human IgG1 CH3 domain. They exhibit superior properties as compared to whole antibodies due to their smaller size and less complex composition, and also as compared to scFvs due to the two antigen-binding domains, for immunotherapy and imaging of various carcinomas including breast cancer. In the current study, efficient production of the recombinant anti-MUC-1 minibody for its dominant format (VH-VL) was obtained in the periplasmic space of the Escherichia coli BL21 (DE3) expression system. The active recombinant protein was successfully purified from soluble fraction. Functional assays presented the in vitro targeting properties and specificity of the expressed anti-MUC-1 HL minibody in the MUC-1 positive cell lines compared to normal cell.

TidsskriftProtein Expression and Purification
StatusUdgivet - nov. 2022

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