Figure 1.
Definition of carboxyl particles and their coupling to proteins, antibodies, peptides, oligonucleotides, etc. – (a) beads contain internal dyes of different intensities; (b) chemistry occurs where the primary amine group of the target molecule (a protein, antibody, or peptide) interacts with the microsphere’s carboxyl groups either directly or indirectly to generate a covalent amide bond.
Figure 2.
Bead array assay description – (a) distribution of the coupled beads with the targeted biomolecule (in this example, an antigen) in the 96-well microtiter plate, followed by the introduction of the experimental serum or plasma; (b) coupled beads bind and immobilize protein-specific antibodies; (c) antibodies that have been immobilized on the beads are differentiated and identified through the utilization of specific secondary antibodies that possess unique labels, regardless of whether they belong to the same or distinct isotypes.
Figure 3.
Executing the analysis – (a) acquisition of the samples using a flow cytometry platform; (b) producing and gathering data and aggregating the outcomes obtained from the various detection antibodies.
Figure 4.
Data analysis obtained from flow cytometry using specialized software – (a) identifying individual sets of beads based on their intensity using two discrimination channels; (b) identification and differentiation of samples that test positive versus negative.