Development and characterization of a human single-chain antibody fragment against claudin-3: a novel therapeutic target in ovarian and uterine carcinomas.

Abstract

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.

• Authors: Chiara Romani, Fabrizio Comper, Elisabetta Bandiera, Antonella Ravaggi, Eliana Bignotti, Renata A Tassi, Sergio Pecorelli, Alessandro D Santin
• Journal: American journal of obstetrics and gynecology (Am J Obstet Gynecol) Vol. 201 Issue 1 Pg. 70.e1-9 (Jul 2009) ISSN: 1097-6868 [Electronic] United States
• PMID: 19426958 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't)

Chemical References

• CLDN3 protein, human
• Claudin-3
• Immunoglobulin Fragments
• Membrane Proteins

Topics

• Carcinoma, Papillary (immunology, metabolism)
• Cell Line, Tumor
• Cell Separation
• Claudin-3
• Enzyme-Linked Immunosorbent Assay
• Female
• Flow Cytometry
• Gene Library
• Humans
• Immunoglobulin Fragments (immunology, metabolism)
• Membrane Proteins (immunology, metabolism)
• Microscopy, Confocal
• Ovarian Neoplasms (immunology, metabolism)
• Self-Sustained Sequence Replication
• Sensitivity and Specificity
• Surface Plasmon Resonance
• Uterine Cervical Neoplasms (immunology, metabolism)