Monday, November 9, 2009
P15
Engineering of antibody fragments from mAb 84, a cytotoxic antibody targeting undifferentiated human embryonic stem cells
Yi-Han Ng, Andre B. Choo, and Victor V.T. Wong. Bioprocessing Technology Institute, A*Star, 20 Biopolis Way, Centros, #06-01, Singapore, 138668, Singapore
The monoclonal antibody, mAb 84, is an IgM which specifically binds and kills undifferentiated human embryonic stem cells (hESC). In cell therapy applications using hESC-derived cells, contaminating undifferentiated hESC can pose a risk of teratoma formation. Thus, mAb 84 can potentially be used for separation and removal of residual undifferentiated hESC from differentiated cell populations. Compared to whole antibodies, antibody fragments may afford better penetration into hESC-derived cell clumps due to their small size. In this study, several antibody fragment constructs of mAb 84 were engineered, including monovalent single-chain variable fragments (scFv 84) and antigen-binding fragments (Fab 84), as well as multivalent fragments of scFv. The antibody fragments were expressed as soluble proteins in Escherichia coli and purified by immobilized metal-ion affinity chromatography or ion-exchange chromatography. All of the antibody fragments retained binding specificity to undifferentiated hESC. Moreover, one of the multivalent fragments was cytotoxic to undifferentiated hESC.
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