Recombinant variants of antibody 138H11 against human gamma-glutamyltransferase for targeting renal cell carcinoma.
The monoconal antibody 138H11 recognizes human renal gamma-glutamyltransferase (GGT), an antigen shown to allow targeting primary and metastatic renal cell carcinoma (RCC). We determined the primary structure of the antigen binding region of mAb 138H11 and generated several different recombinant antibody variants. First, monomeric single-chain Fv antibody fragments, diabodies and triabodies were obtained by constructing linker variants consisting of 18, 10, 8, 5, 3, 2, 1 and zero amino acid residues, resulting in significant differences in yield and molecular architecture. Second, two variants of disulphide bond-stabilised Fv fragments (dsFvs) were generated using two different pairs of complementary framework amino acid positions of VH and VL for disulphide stabilisation. The binding activities of diabodies to human GGT located on its tissue decreased when using shorter linker lengths. The scFv dimer containing a 3 amino acid residue linker peptide and one of the dsFv variants were not functional. Further, the work paves the way for generating new effector constructs and a systematic optimisation of the pharmacokinetics of this tumor targeting antibody by offering variants with a broad range of valencies and molecular masses.
Schmiedl
A
A
Institut Virion\Serion GmbH, Friedrich-Bergius-Ring 19, 97076 Würzburg, Germany.
Zimmermann
J
J
Scherberich
J E
JE
Fischer
P
P
Dübel
S
S
D000595:Amino Acid Sequence; D000906:Antibodies / Q000276:immunology*; D001483:Base Sequence; D015153:Blotting, Western; D002292:Carcinoma, Renal Cell / Q000201:enzymology*; D017931:DNA Primers; D004273:DNA, Neoplasm; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007680:Kidney Neoplasms / Q000201:enzymology*; D008969:Molecular Sequence Data; D011995:Recombination, Genetic; D005723:gamma-Glutamyltransferase / Q000276:immunology*
D000906:Antibodies; D017931:DNA Primers; D004273:DNA, Neoplasm; D005723:gamma-Glutamyltransferase