Binding to human colon cancer cells HCT 116 was analyzed by flow cytometry for purified antibody variants of lgGl-hDR5-01-G56T and lgGl-hDR5-05 with and without the E430G mutation. Single cell suspensions were prepared and binding was analyzed for serial dilution antibody preparation series (range 0.0006 to 10 μg/mL final concentrations in 4- fold dilutions) as described in Example 3. After incubation with the secondary antibody, cells were washed twice, resuspended in 100 μί FACS buffer, and antibody binding was analyzed on a BD LRSFFortessa cell analyzer (BD Biosciences). Bispecific IgGl antibodies were generated by…
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