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

Mona Pourjafar; Michaela Miehe; Rezvan Najafi; Meysam Soleimani; Edzard Spillner

doi:10.1016/j.pep.2022.106148

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

Protein Expr Purif. 2022 Nov:199:106148. doi: 10.1016/j.pep.2022.106148. Epub 2022 Aug 6.

Authors

Mona Pourjafar¹, Michaela Miehe², Rezvan Najafi³, Meysam Soleimani⁴, Edzard Spillner⁵

Affiliations

¹ Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran; Immunological Biotechnology, Department of Biological and Chemical Engineering, Aarhus University, Aarhus, Denmark.
² Immunological Biotechnology, Department of Biological and Chemical Engineering, Aarhus University, Aarhus, Denmark.
³ Research Center for Molecular Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.
⁴ Department of Pharmaceutical Biotechnology, School of Pharmacy, Hamadan University of Medical Sciences, Hamadan, Iran. Electronic address: m.soleymani@umsha.ac.ir.
⁵ Immunological Biotechnology, Department of Biological and Chemical Engineering, Aarhus University, Aarhus, Denmark. Electronic address: e.spillner@bce.au.dk.

PMID: 35940518
DOI: 10.1016/j.pep.2022.106148

Abstract

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 coliBL21 (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.

Keywords: Breast cancer; Engineered antibody; Mammalian expression; Minibody; Mucin-1; Periplasmic expression.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antibodies, Monoclonal*
Antigens, Neoplasm / genetics
Humans
Immunotherapy
Recombinant Proteins / chemistry
Single-Chain Antibodies* / genetics

Substances

Antibodies, Monoclonal
Antigens, Neoplasm
Recombinant Proteins
Single-Chain Antibodies