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).

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.