Abstract
Tribodies are multifunctional recombinant antibody derivatives, which utilize the natural in vivo heterodimerization of the heavy chain (Fd fragment) and light chain (L) of a Fab fragment, to form a scaffold, upon which additional functions can be incorporated, such as additional binders – e.g., scFv binding domains.
Each chain can be extended preferably at the C terminus with an additional scFv binder. The chains are coproduced in mammalian cells, where the host-cell BiP chaperone drives the formation of the heavy chain–light chain heterodimer (Fd:L) – this reaction does not appear to be inhibited by the chain extensions, and leads to a very specific heterodimerization, using molecules abundantly present in serum (non-immunogenic)
These heterodimers are stable, with each of the binders retaining their specific affinities, with the bivalent tribody having higher affinity, and higher activation of T-cell proliferation and cytotoxicity in vivo.
This design allows easy engineering of multispecificity in a single molecule, e.g., bispecific antibodies bivalent for the target and monovalent for effector activation (e.g., for T-cell activation), or trispecific antibodies pur sang.
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References
Arndt KM, Muller KM, Pluckthun A (1998) Factors influencing the dimer to monomer transition of an antibody single-chain Fv fragment. Biochemistry 37(37):12918–12926
Baeuerle PA, Kufer P, Bargou R (2009) BiTE: teaching antibodies to engage T-cells for cancer therapy. Curr Opin Mol Ther 11(1):22–30
Borvak J, Richardson J, Medesan C, Antohe F, Radu C, Simionescu M, Ghetie V, Ward ES (1998) Functional expression of the MHC class I-related receptor, FcRn, in endothelial cells of mice. Int Immunol 10(9):1289–1298
Brissinck J, Demanet C, Moser M, Leo O, Thielemans K (1991) Treatment of mice bearing BCL1 lymphoma with bispecific antibodies. J Immunol 147(11):4019–4026
Gimmi CD, Freeman GJ, Gribben JG, Gray G, Nadler LM (1993) Human T-cell clonal anergy is induced by antigen presentation in the absence of B7 costimulation. Proc Natl Acad Sci USA 90(14):6586–6590
Lee YK, Brewer JW, Hellman R, Hendershot LM (1999) BiP and immunoglobulin light chain cooperate to control the folding of heavy chain and ensure the fidelity of immunoglobulin assembly. Mol Biol Cell 10(7):2209–2219
Schoonjans R, Willems A, Grooten J, Mertens N (2000a) Efficient heterodimerization of recombinant bi- and trispecific antibodies. Bioseparation 9(3):179–183
Schoonjans R, Willems A, Schoonooghe S, Fiers W, Grooten J, Mertens N (2000b) Fab chains as an efficient heterodimerization scaffold for the production of recombinant bispecific and trispecific antibody derivatives. J Immunol 165(12):7050–7057
Schoonjans R, Willems A, Schoonooghe S, Leoen J, Grooten J, Mertens N (2001) A new model for intermediate molecular weight recombinant bispecific and trispecific antibodies by efficient heterodimerization of single chain variable domains through fusion to a Fab-chain. Biomol Eng 17(6):193–202
Schoonooghe S, Kaigorodov V, Zawisza M, Dumolyn C, Haustrate J, Grooten J, Mertens N (2009) Efficient production of human bivalent and trivalent anti-MUC1 Fab-scFv antibodies in Pichia pastoris. BMC Biotechnol 9(1):70
Schoonooghe S, Burvenich I, Vervoort L, De Vos F, Mertens N, Grooten J (2010) PH1-derived bivalent bibodies and trivalent tribodies bind differentially to shed and tumour cell-associated MUC1. Protein Eng Des Sel 23(9):721–728
Smith BJ, Popplewell A, Athwal D, Chapman AP, Heywood S, West SM, Carrington B, Nesbitt A, Lawson AD, Antoniw P, Eddelston A, Suitters A (2001) Prolonged in vivo residence times of antibody fragments associated with albumin. Bioconjug Chem 12(5):750–756
Stebbings R, Findlay L, Edwards C, Eastwood D, Bird C, North D, Mistry Y, Dilger P, Liefooghe E, Cludts I, Fox B, Tarrant G, Robinson J, Meager T, Dolman C, Thorpe SJ, Bristow A, Wadhwa M, Thorpe R, Poole S (2007) “Cytokine storm” in the phase I trial of monoclonal antibody TGN1412: better understanding the causes to improve preclinical testing of immunotherapeutics. J Immunol 179(5):3325–3331
Steffens MG, Boerman OC, Oosterwijk-Wakka JC, Oosterhof GO, Witjes JA, Koenders EB, Oyen WJ, Buijs WC, Debruyne FM, Corstens FH, Oosterwijk E (1997) Targeting of renal cell carcinoma with iodine-131-labeled chimeric monoclonal antibody G250. J Clin Oncol 15(4):1529–1537
Todd PA, Brogden RN (1989) Muromonab CD3. A review of its pharmacology and therapeutic potential. Drugs 37(6):871–899
Willems A, Leoen J, Schoonooghe S, Grooten J, Mertens N (2003) Optimizing expression and purification from cell culture medium of trispecific recombinant antibody derivatives. J Chromatogr B 786(1–2):161–176
Willems A, Schoonooghe S, Eeckhout D, De Jaegher G, Grooten J, Mertens N (2005) CD3 × CD28 cross-interacting bispecific antibodies improve tumor cell dependent T-cell activation. Cancer Immunol Immunother 54:1059–1071
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Mertens, N. (2011). Tribodies: Fab–scFv Fusion Proteins as a Platform to Create Multifunctional Pharmaceuticals. In: Kontermann, R. (eds) Bispecific Antibodies. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-20910-9_8
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DOI: https://doi.org/10.1007/978-3-642-20910-9_8
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