The first constant domain (C(H)1 and C(L)) of an antibody used as heterodimerization domain for bispecific miniantibodies.
Bispecific miniantibodies were constructed by genetically fusing the C(H)1 domain of an IgG1 to the C-terminus of a single-chain Fv fragment (scFv-425), specific for the EGF receptor, and fusing the C(L) domain of a kappa light chain to the C-terminus of a scFv specific for CD2 (scFv-M1). An efficient dicistronic gene arrangement for functional expression in Escherichia coli was constructed. Immunoblots demonstrated correct domain assembly and the formation of the natural C(H)1-C(L) disulfide bridge. Gel filtration confirmed the correct size, sandwich ELISAs demonstrated bispecific functionality, and SPR biosensor measurements determined binding to EGF-R in comparison to bivalent constructs. Bispecific anti-EGF-R/anti-CD2 miniantibodies are candidates for the immunotherapy of cancer.
Müller
K M
KM
Biochemisches Institut, Universität Zürich, Switzerland.
Arndt
K M
KM
Strittmatter
W
W
Plückthun
A
A
D000595:Amino Acid Sequence; D018033:Antibodies, Bispecific / Q000096:biosynthesis / Q000737:chemistry*; D000918:Antibody Specificity; D001483:Base Sequence; D001666:Binding Sites, Antibody; D018801:CD2 Antigens / Q000276:immunology*; D003001:Cloning, Molecular; D019281:Dimerization; D066246:ErbB Receptors / Q000276:immunology*; D004926:Escherichia coli; D005803:Genes, Immunoglobulin; D006801:Humans; D007127:Immunoglobulin Constant Regions / Q000096:biosynthesis / Q000737:chemistry*; D007074:Immunoglobulin G / Q000096:biosynthesis / Q000737:chemistry*; D007143:Immunoglobulin Heavy Chains / Q000096:biosynthesis / Q000737:chemistry*; D007145:Immunoglobulin kappa-Chains / Q000096:biosynthesis / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D015183:Restriction Mapping
D018033:Antibodies, Bispecific; D018801:CD2 Antigens; D007127:Immunoglobulin Constant Regions; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007145:Immunoglobulin kappa-Chains; D011994:Recombinant Proteins; D066246:ErbB Receptors