The assembly of single domain antibodies into bispecific decavalent molecules.
Bispecific antibodies present unique opportunities in terms of new applications for engineered antibodies. However, designing ideal bispecific antibodies remains a challenge. Here we describe a novel bispecific antibody model in which five single domain antibodies (sdAbs) are fused via a linker sequence to the N-terminus of the verotoxin B (VTB) subunit, a pentamerization domain, and five sdAbs are fused via a linker sequence to the VTB C-terminus. Fifteen such decavalent bispecific molecules, termed decabodies, were constructed and characterized for the purpose of identifying an optimal decabody design. One of the fifteen molecules existed in a non-aggregated decavalent form. In conjunction with the isolation of sdAbs with the desired specificities from non-immune phage display libraries, the decabody strategy provides a means of generating high avidity bispecific antibody reagents, with good physical properties, relatively quickly.
Stone
Emily
E
Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, Canada K1A 0R6.
Hirama
Tomoko
T
Tanha
Jamshid
J
Tong-Sevinc
Hong
H
Li
Shenghua
S
MacKenzie
C Roger
CR
Zhang
Jianbing
J
D000595:Amino Acid Sequence; D000818:Animals; D018033:Antibodies, Bispecific / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D000915:Antibody Affinity / Q000276:immunology; D000937:Antigen-Antibody Reactions / Q000276:immunology; D002161:Camelids, New World; D002850:Chromatography, Gel; D005822:Genetic Vectors / Q000235:genetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D044162:Parathyroid Hormone-Related Protein / Q000276:immunology; D010446:Peptide Fragments / Q000276:immunology; D021122:Protein Subunits / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000276:immunology*; D022603:Shiga Toxins / Q000235:genetics*; D020349:Surface Plasmon Resonance
D018033:Antibodies, Bispecific; D044162:Parathyroid Hormone-Related Protein; D010446:Peptide Fragments; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D022603:Shiga Toxins