Enhanced ADCC Activity of Affinity Maturated and Fc-engineered Mini-antibodies Directed Against the AML Stem Cell Antigen CD96

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2012 Aug 11

PMID 22879978

Citations 12

Authors

Sahar Mohseni Nodehi

Roland Repp

Christian Kellner

Joachim Brautigam

Matthias Staudinger

Natalie Schub

Matthias Peipp

Martin Gramatzki

Andreas Humpe

Affiliations

Soon will be listed here.

Abstract

CD96, a cell surface antigen recently described to be preferentially expressed on acute myeloid leukemia (AML) leukemic stem cells (LSC) may represent an interesting target structure for the development of antibody-based therapeutic approaches. The v-regions from the CD96-specific hybridoma TH-111 were isolated and used to generate a CD96-specific single chain fragment of the variable regions (scFv). An affinity maturated variant resulting in 4-fold enhanced CD96-binding was generated by random mutagenesis and stringent selection using phage display. The affinity maturated scFv CD96-S32F was used to generate bivalent mini-antibodies by genetically fusing an IgG1 wild type Fc region or a variant with enhanced CD16a binding. Antibody dependent cell-mediated cytotoxicity (ADCC) experiments revealed that Fc engineering was essential to trigger significant effector cell-mediated lysis when the wild type scFv was used. The mini-antibody variant generated by fusing the affinity-maturated scFv with the optimized Fc variant demonstrated the highest ADCC activity (2.3-fold enhancement in efficacy). In conclusion, our data provide proof of concept that CD96 could serve as a target structure for effector cell-mediated lysis and demonstrate that both enhancing affinity for CD96 and for CD16a resulted in mini-antibodies with the highest cytolytic potential.

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