Vorsetuzumab

A phagocytosis functional assay measures the ability of phagocytic cells, such as neutrophils, macrophages, monocytes, or microglia/macrophages, to bind and internalize particulate targets including bacteria, yeast particles, beads, or myelin particles. Fluorescent flow-cytometry assays detect target uptake as fluorescence associated with gated phagocytes, while pH-sensitive dyes such as pHrodo increase signal in acidic phagosomal compartments and therefore preferentially report internalized particles rather than particles remaining outside the cell. Microscopy or high-content imaging can be used to confirm intracellular localization and, in some protocols, to follow uptake kinetics.

Recombinant antibody expression produces antibody chains from cloned antibody genes in mammalian cells, commonly HEK293-derived cells, so that secreted IgG or Fc-fusion antibodies can be collected from culture supernatant. Protein A/G purification is affinity chromatography in which immobilized Protein A, Protein G, or Protein A/G binds the antibody Fc region; nonbound culture components are washed away, and bound antibody is recovered by changing buffer conditions, commonly acidic elution followed by neutralization.