The recognition of human HER2 by biosimilar candidate to trastuzumab contained in cell culture supernatant was evaluated by flow cytometry. Tumoral cell lines overexpressing HER2 molecule: SKBR3 and SKOV3 were used. Ramos cells (CD20 +) were also used as a negative control of HER2 expression and recognition. Cells were stained with 10 μg/mL of produced antibody contained in cell culture supernatants or commercial trastuzumab (anti-HER2; Roche, Argentine) for 30 min at 4 °C. A biosimilar candidate to rituximab (anti-CD20) antibody contained in cell culture supernatant (10 μg/mL) was added as isotype-matched control. Cells were washed with PBS and the binding of the antibodies was detected by incubation with a FITC-labeled rabbit anti-human IgG antibody (F0315, Dako, USA) for 30 min at 4 °C. Cells were analyzed on a Sysmex flow cytometer (Germany) and data were processed with FlowJo 7.6.1 software (Tree Star Inc., USA).

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