TY - JOUR
T1 - Generation and characterization of monospecific and bispecific hexavalent trimerbodies
AU - Blanco-Toribio, Ana
AU - Sainz-Pastor, Noelia
AU - Álvarez-Cienfuegos, Ana
AU - Merino, Nekane
AU - Cuesta, Ángel M.
AU - Sánchez-Martín, David
AU - Bonet, Jaume
AU - Santos-Valle, Patricia
AU - Sanz, Laura
AU - Oliva, Baldo
AU - Blanco, Francisco J.
AU - Álvarez-Vallina, Luis
N1 - Funding Information:
This study was supported by grants from Ministerio de Ciencia e Innovación (BIO2008–03233), Ministerio de Economía y Competitividad (BIO2011–22738), and Comunidad de Madrid (S-BIO-0236–2006 and S2010/BMD-2312) to L.A-V.; from Fondo de Investigación Sanitaria/Instituto de Salud Carlos III (PI08/90856 and PS09/00227) to L.S.; Ministerio de Ciencia e Innovación (BIO2008–00205) to B.O., and CTQ2011–28680 to F.J.B. A.B-T. and A.A-C. were supported by Programa Torres Quevedo from Ministerio de Economía y Competitividad, cofounded by the European Social Fund (PTQ-09–01–01089 and PTQ11–04604, respectively). P.S-V. was supported by a Fundación de Investigación Biomédica-Hospital Universitario Puerta de Hierro training grant. The authors declare no conflict of interest related to this work.
PY - 2013/1
Y1 - 2013/1
N2 - Here, we describe a new class of multivalent and multispecific antibody-based reagents for therapy. The molecules, termed "trimerbodies, " use a modified version of the N-terminal trimerization region of human collagen XVIII noncollagenous 1 domain flanked by two flexible linkers as trimerizing scaffold. By fusing single-chain variable fragments (scFv) with the same or different specificity to both N-and C-terminus of the trimerizing scaffold domain, we produced monospecific or bispecific hexavalent molecules that were efficiently secreted as soluble proteins by transfected mammalian cells. A bispecific anti-laminin x anti-CD3 N-/C-trimerbody was found to be trimeric in solution, very efficient at recognizing purified plastic-immobilized laminin and CD3 expressed at the surface of T cells, and remarkably stable in human serum. The bispecificity was further demonstrated in T cell activation studies. In the presence of laminin-rich substrate, the bispecific anti-laminin x anti-CD3 N-/C-trimerbody stimulated a high percentage of human T cells to express surface activation markers. These results suggest that the trimerbody platform offers promising opportunities for the development of the next-generation therapeutic antibodies, i.e., multivalent and bispecific molecules with a format optimized for the desired pharmacokinetics and adapted to the pathological context.
AB - Here, we describe a new class of multivalent and multispecific antibody-based reagents for therapy. The molecules, termed "trimerbodies, " use a modified version of the N-terminal trimerization region of human collagen XVIII noncollagenous 1 domain flanked by two flexible linkers as trimerizing scaffold. By fusing single-chain variable fragments (scFv) with the same or different specificity to both N-and C-terminus of the trimerizing scaffold domain, we produced monospecific or bispecific hexavalent molecules that were efficiently secreted as soluble proteins by transfected mammalian cells. A bispecific anti-laminin x anti-CD3 N-/C-trimerbody was found to be trimeric in solution, very efficient at recognizing purified plastic-immobilized laminin and CD3 expressed at the surface of T cells, and remarkably stable in human serum. The bispecificity was further demonstrated in T cell activation studies. In the presence of laminin-rich substrate, the bispecific anti-laminin x anti-CD3 N-/C-trimerbody stimulated a high percentage of human T cells to express surface activation markers. These results suggest that the trimerbody platform offers promising opportunities for the development of the next-generation therapeutic antibodies, i.e., multivalent and bispecific molecules with a format optimized for the desired pharmacokinetics and adapted to the pathological context.
KW - Antibody engineering
KW - Bispecific antibodies
KW - Collagen
KW - Multivalent antibodies
KW - Trimerbody
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U2 - 10.4161/mabs.22698
DO - 10.4161/mabs.22698
M3 - Article
C2 - 23221741
AN - SCOPUS:84872854587
SN - 1942-0862
VL - 5
SP - 70
EP - 79
JO - mAbs
JF - mAbs
IS - 1
ER -