Human antibody derivatives against the fibroblast activation protein for tumor stroma targeting of carcinomas.
The fibroblast activation protein (FAP) is selectively expressed on activated fibroblasts of the tumor stroma on more than 90% of lung, breast and colon carcinomas. The high prevalence and abundance of FAP(+) stroma make it a promising target for in vivo diagnosis and therapy of a variety of carcinomas. We describe the humanization of the murine FAP-specific MAb, F19, which has already been clinically used for in vivo diagnostic purposes. Using phage display technology and human V-repertoires, VL and VH regions of F19 were replaced by analogous human V-regions while retaining the original HCDR3 sequence in order to maintain F19 epitope specificity. The resulting human single-chain fragments of immunoglobulin variable regions (scFv 34, scFv 18) showed affinities of 6 nM on cell membrane-bound FAP. scFv 34 was expressed as a bivalent minibody (Mb 34). The antigen-binding characteristics of Mb 34 were comparable to the parental and a complementarity-determining region (CDR)-grafted version of F19. This was revealed by binding competition studies, FACS analyses and immunohistochemistry on various tumor samples including breast, colon and lung carcinomas. Importantly, compared with the CDR-grafted humanized scFv version of F19, the V-regions of the selected human scFv 34 showed sequence identity with the parental antibody (Ab) only over the short, 15-amino acid long HCDR3. Thus, a largely reduced xenoantigenic potential is expected. These human Ab derivatives are suitable to develop novel therapeutic concepts with broad applicability for a wide variety of histological carcinomas based on tumor stroma targeting.
Mersmann
M
M
Institut für Zellbiologie und Immunologie, Universität Stuttgart, Allmandring 32, D-70569 Stuttgart, Germany.
Schmidt
A
A
Rippmann
J F
JF
Wüest
T
T
Brocks
B
B
Rettig
W J
WJ
Garin-Chesa
P
P
Pfizenmaier
K
K
Moosmayer
D
D
D000595:Amino Acid Sequence; D000818:Animals; D018033:Antibodies, Bispecific / Q000235:genetics / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D000951:Antigens, Neoplasm / Q000276:immunology* / Q000378:metabolism; D014408:Biomarkers, Tumor; D002277:Carcinoma / Q000276:immunology* / Q000378:metabolism; D010450:Endopeptidases; D000939:Epitopes / Q000276:immunology; D016326:Extracellular Matrix Proteins / Q000276:immunology / Q000378:metabolism; D018093:Gelatinases; D006133:Growth Substances / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology; D008565:Membrane Proteins; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D011993:Recombinant Fusion Proteins / Q000276:immunology; D017386:Sequence Homology, Amino Acid; D012697:Serine Endopeptidases / Q000276:immunology* / Q000378:metabolism; D013696:Temperature; D000099167:Fibroblast Activation Protein Alpha
D018033:Antibodies, Bispecific; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D010450:Endopeptidases; D000939:Epitopes; D016326:Extracellular Matrix Proteins; D018093:Gelatinases; D006133:Growth Substances; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D008565:Membrane Proteins; D019151:Peptide Library; D011993:Recombinant Fusion Proteins; D012697:Serine Endopeptidases; D000099167:Fibroblast Activation Protein Alpha