Antigen binding and stability properties of non-covalently linked anti-CD22 single-chain Fv dimers.
By varying linker length and domain orientation three multivalent derivatives of a monovalent anti-CD22 single-chain fragment variable (scFv) antibody were generated. Shortening the linker of the V(H)-V(L) oriented scFv to 5 or 0 residues resulted in the formation of diabodies or a mixture of tetramers and trimers, respectively. Unexpectedly, a V(L)-0-V(H) scFv assembled to homogenous dimers, remained substantially more stable than the V(H)-5-V(L) diabody when incubated in human serum at 37 degrees C, and retained its dimeric state when concentrated up to 4 mg/ml. These properties suggest the V(L)-0-V(H) scFv could become an attractive vehicle for the selective delivery of multiple effector molecules to CD22(+) tumor cells.
Arndt
Michaela A E
MA
SAIC, National Cancer Institute at Frederick, Frederick, MD 21702, USA. michaela.arndt@medizin.uni-essen.de
Krauss
Jürgen
J
Rybak
Susanna M
SM
D000595:Amino Acid Sequence; D018033:Antibodies, Bispecific / Q000378:metabolism; D000915:Antibody Affinity / Q000276:immunology*; D015703:Antigens, CD / Q000378:metabolism*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000378:metabolism*; D001666:Binding Sites, Antibody; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002460:Cell Line; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D019281:Dimerization; D004926:Escherichia coli / Q000235:genetics; D005434:Flow Cytometry; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000737:chemistry* / Q000276:immunology; D019169:Jurkat Cells; D037102:Lectins / Q000378:metabolism*; D016393:Lymphoma, B-Cell / Q000276:immunology; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011947:Receptors, Antigen, B-Cell / Q000378:metabolism*; D051918:Sialic Acid Binding Ig-like Lectin 2; D013696:Temperature
D018033:Antibodies, Bispecific; D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; C497867:CD22 protein, human; D015815:Cell Adhesion Molecules; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D037102:Lectins; D011947:Receptors, Antigen, B-Cell; D051918:Sialic Acid Binding Ig-like Lectin 2