Figure 7.

Purification of monomeric Fab fragment T1-1 and inhibition of HeLa cell growth in the Alamar Blue® surrogate apoptosis assay. The relative molecular mass of the purified T1-1 Fab as determined by size exclusion chromatography is shown in (A). For calibration, the elution peaks for four standard proteins (alcohol dehydrogenase, Mw 150 kDa; bovine serum albumin, Mw 66 kDa; carbonic anhydrase, Mw 29 kDa and cytochrome C, Mw 12 kDa) are shown as crosses. A single peak is seen at the predicted Mw for a monomeric Fab fragment (∼50 kDa), with no evidence of higher Mw peaks. The effect of T1-1 Fab on HeLa cell viability is shown in (B). Cell survival, expressed as % HeLa cell proliferation, is shown following incubation with increasing concentrations of T1-1 Fab (filled squares, red) and an irrelevant Fab (filled diamonds, grey).