OBJECTIVE: The purpose of this study was to develop and characterize a human antibody in a single-chain antibody fragment format (scFv) that is directed specifically against claudin-3 (CLDN3). STUDY DESIGN: The synthetic ETH-2 Gold human antibody phage display library was used to select scFv specific against CLDN3. scFv binding properties were analyzed by surface plasmon resonance; specificity was confirmed with enzyme-linked immunosorbent assay, immunofluorescence, and flow cytometry on a panel of ovarian and uterine serous carcinoma cell lines. RESULTS: Surface plasmon resonance studies indicated scFv H6 to be the clone with the highest affinity against CLDN3 (K(D) of 23.60 nmol/L). scFv H6 efficiently stained CLDN3-expressing cells and recognized its epitope in enzyme-linked immunosorbent assay that was performed with uterine serous papillary carcinoma native protein extract, which suggested that a conformational epitope is recognized by this antibody. Cell surface immunofluorescence with laser scanning confocal microscopy confirmed the specific binding to the native membrane CLDN3. CONCLUSION: scFv H6 may represent a novel antitumor agent against chemotherapy-resistant ovarian and serous papillary carcinomas and other human malignancies that overexpress CLDN3.
OBJECTIVE: The purpose of this study was to develop and characterize a human antibody in a single-chain antibody fragment format (scFv) that is directed specifically against claudin-3 (CLDN3). STUDY DESIGN: The synthetic ETH-2 Gold human antibody phage display library was used to select scFv specific against CLDN3. scFv binding properties were analyzed by surface plasmon resonance; specificity was confirmed with enzyme-linked immunosorbent assay, immunofluorescence, and flow cytometry on a panel of ovarian and uterine serous carcinoma cell lines. RESULTS: Surface plasmon resonance studies indicated scFv H6 to be the clone with the highest affinity against CLDN3 (K(D) of 23.60 nmol/L). scFv H6 efficiently stained CLDN3-expressing cells and recognized its epitope in enzyme-linked immunosorbent assay that was performed with uterine serous papillary carcinoma native protein extract, which suggested that a conformational epitope is recognized by this antibody. Cell surface immunofluorescence with laser scanning confocal microscopy confirmed the specific binding to the native membrane CLDN3. CONCLUSION:scFv H6 may represent a novel antitumor agent against chemotherapy-resistant ovarian and serous papillary carcinomas and other humanmalignancies that overexpress CLDN3.
Authors: G P Adams; R Schier; A M McCall; H H Simmons; E M Horak; R K Alpaugh; J D Marks; L M Weiner Journal: Cancer Res Date: 2001-06-15 Impact factor: 12.701
Authors: F Turatti; D Mezzanzanica; E Nardini; E Luison; L Maffioli; E Bambardieri; C de Lalla; S Canevari; M Figini Journal: Cancer Immunol Immunother Date: 2001-02 Impact factor: 6.968
Authors: P Michl; M Buchholz; M Rolke; S Kunsch; M Löhr; B McClane; S Tsukita; G Leder; G Adler; T M Gress Journal: Gastroenterology Date: 2001-09 Impact factor: 22.682
Authors: Emiliano Cocco; Francesca Casagrande; Stefania Bellone; Christine E Richter; Marta Bellone; Paola Todeschini; Jennie C Holmberg; Han Hsuan Fu; Michele K Montagna; Gil Mor; Peter E Schwartz; Dan Arin-Silasi; Masoud Azoudi; Thomas J Rutherford; Maysa Abu-Khalaf; Sergio Pecorelli; Alessandro D Santin Journal: BMC Cancer Date: 2010-07-02 Impact factor: 4.430