Dimeric assembly of antibody fragments and other therapeutic molecules can result in increased binding and improved bioactivity. Here, we investigated the use of the IgM heavy chain domain 2 (MHD2) as covalently linked homodimerization module. Fusion of single-chain fragment variable (scFv) molecules directed against epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 to the N- and/or C-terminus of the MHD2, respectively, resulted in molecules with single or dual specificity for tumor cells. Bispecific tetravalent molecules were further generated by fusing a bispecific single-chain diabody directed against EGFR and epithelial cell adhesion molecule to the N-terminus of the MHD2. By combining an anti-EGFR scFv with a single-chain derivative of tumor necrosis factor, a tetravalent bifunctional fusion protein was produced. This fusion protein exhibited improved TNF activity, also mimicking the membrane-bound form of TNF, as shown by the activation of TNFR2-mediated cell killing. Furthermore, the scFv moiety allowed for an antigen-dependent delivery of TNF to EGFR-positive cells and an improved stimulatory TNF action on these cells. Thus, we established the MHD2 as a versatile module for the generation of bispecific and bifunctional fusion proteins.
%0 Journal Article
%1 Seifert2012
%A Seifert, Oliver
%A Plappert, Aline
%A Heidel, Nadine
%A Fellermeier, Sina
%A Messerschmidt, Sylvia K E
%A Richter, Fabian
%A Kontermann, Roland E.
%D 2012
%J Protein Engineering, Design and Selection
%K 2012 izi kontermann
%N 10
%P 603--612
%R 10.1093/protein/gzs059
%T The IgM CH2 domain as covalently linked homodimerization module for the generation of fusion proteins with dual specificity
%U http://www.ncbi.nlm.nih.gov/pubmed/22988132
%V 25
%X Dimeric assembly of antibody fragments and other therapeutic molecules can result in increased binding and improved bioactivity. Here, we investigated the use of the IgM heavy chain domain 2 (MHD2) as covalently linked homodimerization module. Fusion of single-chain fragment variable (scFv) molecules directed against epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 to the N- and/or C-terminus of the MHD2, respectively, resulted in molecules with single or dual specificity for tumor cells. Bispecific tetravalent molecules were further generated by fusing a bispecific single-chain diabody directed against EGFR and epithelial cell adhesion molecule to the N-terminus of the MHD2. By combining an anti-EGFR scFv with a single-chain derivative of tumor necrosis factor, a tetravalent bifunctional fusion protein was produced. This fusion protein exhibited improved TNF activity, also mimicking the membrane-bound form of TNF, as shown by the activation of TNFR2-mediated cell killing. Furthermore, the scFv moiety allowed for an antigen-dependent delivery of TNF to EGFR-positive cells and an improved stimulatory TNF action on these cells. Thus, we established the MHD2 as a versatile module for the generation of bispecific and bifunctional fusion proteins.
%@ 1741-0134 (Electronic)$\backslash$r1741-0126 (Linking)
@article{Seifert2012,
abstract = {Dimeric assembly of antibody fragments and other therapeutic molecules can result in increased binding and improved bioactivity. Here, we investigated the use of the IgM heavy chain domain 2 (MHD2) as covalently linked homodimerization module. Fusion of single-chain fragment variable (scFv) molecules directed against epidermal growth factor receptor (EGFR) and human epidermal growth factor receptor 2 to the N- and/or C-terminus of the MHD2, respectively, resulted in molecules with single or dual specificity for tumor cells. Bispecific tetravalent molecules were further generated by fusing a bispecific single-chain diabody directed against EGFR and epithelial cell adhesion molecule to the N-terminus of the MHD2. By combining an anti-EGFR scFv with a single-chain derivative of tumor necrosis factor, a tetravalent bifunctional fusion protein was produced. This fusion protein exhibited improved TNF activity, also mimicking the membrane-bound form of TNF, as shown by the activation of TNFR2-mediated cell killing. Furthermore, the scFv moiety allowed for an antigen-dependent delivery of TNF to EGFR-positive cells and an improved stimulatory TNF action on these cells. Thus, we established the MHD2 as a versatile module for the generation of bispecific and bifunctional fusion proteins.},
added-at = {2023-06-29T13:07:55.000+0200},
author = {Seifert, Oliver and Plappert, Aline and Heidel, Nadine and Fellermeier, Sina and Messerschmidt, Sylvia K E and Richter, Fabian and Kontermann, Roland E.},
biburl = {https://puma.ub.uni-stuttgart.de/bibtex/213019e84da8e964a2efaa913253e9230/fabian},
doi = {10.1093/protein/gzs059},
interhash = {a2066fd3c673be45c4181140e32b68d4},
intrahash = {13019e84da8e964a2efaa913253e9230},
isbn = {1741-0134 (Electronic)$\backslash$r1741-0126 (Linking)},
issn = {17410126},
journal = {Protein Engineering, Design and Selection},
keywords = {2012 izi kontermann},
month = oct,
number = 10,
pages = {603--612},
pmid = {22988132},
timestamp = {2023-06-29T13:07:55.000+0200},
title = {{The IgM CH2 domain as covalently linked homodimerization module for the generation of fusion proteins with dual specificity}},
url = {http://www.ncbi.nlm.nih.gov/pubmed/22988132},
volume = 25,
year = 2012
}
