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

Mona Pourjafar, Michaela Miehe, Rezvan Najafi, Meysam Soleimani^*, Edzard Spillner

^*Corresponding author for this work

Department of Biological and Chemical Engineering - Industrial Biotechnology - Gustav Wieds Vej 10D
Department of Biological and Chemical Engineering - Medical Biotechnology - Gustav Wieds Vej 10D

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

3 Citations (Scopus)

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 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.

Original language	English
Article number	106148
Journal	Protein Expression and Purification
Volume	199
ISSN	1046-5928
DOIs	https://doi.org/10.1016/j.pep.2022.106148
Publication status	Published - Nov 2022

Keywords

Breast cancer
Engineered antibody
Mammalian expression
Minibody
Mucin-1
Periplasmic expression

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10.1016/j.pep.2022.106148

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

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 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.",

keywords = "Breast cancer, Engineered antibody, Mammalian expression, Minibody, Mucin-1, Periplasmic expression",

author = "Mona Pourjafar and Michaela Miehe and Rezvan Najafi and Meysam Soleimani and Edzard Spillner",

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year = "2022",

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doi = "10.1016/j.pep.2022.106148",

language = "English",

volume = "199",

journal = "Protein Expression and Purification",

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Construction, expression and functional characterization of an engineered antibody against tumor antigen MUC-1C. / Pourjafar, Mona; Miehe, Michaela; Najafi, Rezvan et al.
In: Protein Expression and Purification, Vol. 199, 106148, 11.2022.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - Construction, expression and functional characterization of an engineered antibody against tumor antigen MUC-1C

AU - Pourjafar, Mona

AU - Miehe, Michaela

AU - Najafi, Rezvan

AU - Soleimani, Meysam

AU - Spillner, Edzard

PY - 2022/11

Y1 - 2022/11

N2 - 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.

AB - 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.

KW - Breast cancer

KW - Engineered antibody

KW - Mammalian expression

KW - Minibody

KW - Mucin-1

KW - Periplasmic expression

U2 - 10.1016/j.pep.2022.106148

DO - 10.1016/j.pep.2022.106148

M3 - Journal article

C2 - 35940518

AN - SCOPUS:85135701459

SN - 1046-5928

VL - 199

JO - Protein Expression and Purification

JF - Protein Expression and Purification

M1 - 106148

ER -

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

Abstract

Keywords

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