Anti-GPVI (HY101) activates platelets in a multicolor flow cytometry panel.

Area of Science:

• Immunology
• Hematology
• Biotechnology

Background:

• Platelet membrane markers, including GPVI (glycoprotein VI), are crucial for identifying platelet subpopulations.
• Multicolor flow cytometry enables simultaneous assessment of multiple platelet markers in whole blood.
• Accurate detection of platelet subgroups relies on the stability of cell surface markers during analysis.

Purpose of the Study:

• To report an unexpected platelet activation issue associated with anti-GPVI antibody clone HY101.
• To investigate the impact of this antibody on platelet activation marker expression in flow cytometry.
• To validate findings using complementary platelet aggregation assays.

Main Methods:

• Whole blood multicolor flow cytometry panel including anti-GPVI (clone HY101) and other platelet markers.
• Assessment of platelet activation markers: CD62P (P-selectin), PAC-1 (activated GPIIb/IIIa), CD63, and CD107a (LAMP-1).
• Platelet aggregation studies as a confirmatory method.

Main Results:

• Addition of anti-GPVI antibody clone HY101 induced significant platelet activation.
• Increased expression of CD62P, PAC-1, CD63, and CD107a was observed in flow cytometry analysis.
• Platelet aggregation studies confirmed the activation triggered by the anti-GPVI antibody.

Conclusions:

• The anti-GPVI antibody clone HY101 exhibits intrinsic platelet-activating properties.
• This activation artifact complicates the accurate assessment of platelet subgroups using flow cytometry.
• Researchers should be aware of this potential issue when using anti-GPVI clone HY101 in flow cytometry panels.