A THP-1 antibody-dependent cellular phagocytosis (ADCP) assay was performed as previously described (55 (link)). Briefly, recombinant HIV proteins were conjugated onto 1-μm fluorescent neutravidin beads (Invitrogen). Excess antigen was removed by washing pelleted beads, which were then incubated with purified IgG antibody samples (100 μg/ml) for 2 h at 37°C. Following opsonization, THP-1 cells were added to the bead-antibody mix and incubated overnight, followed by fixation and analysis of bead uptake by flow cytometry.
Muenchhoff M., Chung A.W., Roider J., Dugast A.S., Richardson S., Kløverpris H., Leslie A., Ndung’u T., Moore P., Alter G, & Goulder P.J. (2020). Distinct Immunoglobulin Fc Glycosylation Patterns Are Associated with Disease Nonprogression and Broadly Neutralizing Antibody Responses in Children with HIV Infection. mSphere, 5(6), e00880-20.
Other organizations :
Ludwig-Maximilians-Universität München, German Center for Infection Research, University of Oxford, University of KwaZulu-Natal, Peter Doherty Institute, University of Melbourne, Africa Health Research Institute, Rubius Therapeutics (United States), University of the Witwatersrand, University of Copenhagen, University College London, Ragon Institute of MGH, MIT and Harvard, Max Planck Institute for Infection Biology
Recombinant HIV proteins conjugated onto 1-μm fluorescent neutravidin beads
Incubation of beads with purified IgG antibody samples (100 μg/ml) for 2 h at 37°C
dependent variables
Bead uptake by THP-1 cells measured by flow cytometry
control variables
Incubation conditions (2 h at 37°C)
controls
Positive control: Not specified
Negative control: Not specified
Annotations
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