THP-1 Phagocytosis Assay for Antibody-Antigen Complexes

THP-1 cellular phagocytosis assay was performed as previously described [15 (link)]. Briefly, biotinylated antigens were bound to FluoSphere NeutrAvidin beads (Thermo Fisher Scientific; Waltham, MA, USA). To form immune-complexes, antigen-coated beads were incubated with 10 µL of 1:100 diluted serum samples. THP-1 monocytes were added to the bead mixture and incubated at 37 °C. After 16 h incubation, cells were fixed with 4% paraformaldehyde and analyzed on an iQue analyzer (Intellicyt; Albuquerque, NM, USA).

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Burns M.D., Boribong B.P., Bartsch Y.C., Loiselle M., St. Denis K.J., Sheehan M.L., Chen J.W., Davis J.P., Lima R., Edlow A.G., Fasano A., Balazs A.B., Alter G, & Yonker L.M. (2022). Durability and Cross-Reactivity of SARS-CoV-2 mRNA Vaccine in Adolescent Children. Vaccines, 10(4), 492.

Publication 2022

FluoSphere NeutrAvidin beads iQue analyzer

Antigen Assay Cells Immune complexes Monocytes Neutravidin Paraformaldehyde Phagocytosis Serum Thp 1 cellular

Corresponding Organization :

Other organizations : Massachusetts General Hospital, Ragon Institute of MGH, MIT and Harvard

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Variable analysis

independent variables

Biotinylated antigens bound to FluoSphere NeutrAvidin beads
Serum samples diluted 1:100

dependent variables

Phagocytosis of immune-complexes by THP-1 monocytes

control variables

Incubation temperature of 37 °C
Incubation time of 16 h

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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