Simultaneous and Independent Dual Site-Specific Self-Labeling of Recombinant Antibodies.

Abstract

Antibody-based diagnostic and therapeutic reagents armed with effector molecules such as dyes and drugs offer hope in the battle against cancer. Several site-specific conjugation methods have been developed to equip antibodies with such effector molecules, but they tend to be expensive and involve multiple reaction steps. The conjugation of two different effector molecules to a single antibody also remains a major challenge. Here we describe a simple, controlled, and robust method for the dual site-specific conjugation of an antibody with two effector molecules in a single-pot reaction using the self-labeling SNAP and CLIP protein tags. We verified the principle of the method by labeling an epidermal growth factor receptor (EGFR)-specific single-chain antibody fragment (scFv-425) simultaneously with IRDye700 and Alexa-Fluor647. This dual-labeled antibody bound to EGFR+ ovarian cancer cell lines and tissue samples with high specificity, and its phototherapeutic efficacy was confirmed by the selective killing of EGFR+ cells in vitro.

• Authors: Carolin Wollschlaeger, Ivo Meinhold-Heerlein, Xiaojing Cong, Karen Bräutigam, Stefano Di Fiore, Felix Zeppernick, Torsten Klockenbring, Elmar Stickeler, Stefan Barth, Ahmad Fawzi Hussain
• Journal: Bioconjugate chemistry (Bioconjug Chem) Vol. 29 Issue 11 Pg. 3586-3594 (11 21 2018) ISSN: 1520-4812 [Electronic] United States
• PMID: 30289242 (Publication Type: Journal Article)

Chemical References

• Coloring Agents
• Immunoconjugates
• Recombinant Fusion Proteins
• Single-Chain Antibodies
• scFv 425
• ErbB Receptors

Topics

• Cell Line, Tumor
• Coloring Agents (chemistry)
• ErbB Receptors (immunology, metabolism)
• Female
• Flow Cytometry
• Humans
• Immunoconjugates (chemistry)
• Microscopy, Confocal
• Ovarian Neoplasms (pathology)
• Protein Binding
• Protein Conformation
• Recombinant Fusion Proteins (chemistry, immunology)
• Single-Chain Antibodies (chemistry, immunology)