Methods for Detecting Antibody Self-Association
AssigneeAmgen Inc.
InventorsYuqing Shen, Emma Montserrat Pelegri-O'Day
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Abstract
The disclosure provides for methods and systems for detecting antibody protein product self-association comprising inducing co-agglutination of nanoparticles and solid supports to capture the nanoparticles, each coated with a ligand specific for immunoglobulins, and utilizing a fluidic device to detect co-agglutination and antibody protein product self-association.
Claims21
A method of detecting antibody protein product association comprising
- a) contacting a sample comprising an antibody protein product with nanoparticles coated with a ligand for an antibody protein product, wherein the contacting occurs in a fluidic device under conditions that allow antibody protein products to self-associate, thereby forming clusters of nanoparticles,
- b) contacting the clusters of nanoparticles with a solid support, wherein the solid support is coated with a ligand for the antibody protein product of (a), and
- c) detecting a change in optical signal of the nanoparticles and/or the solid support, wherein a change in optical signal indicates antibody protein product association.
The method of any of the preceding claims, wherein the antibody protein product comprises or consists of a large peptide, antibody, antibody fragment, antibody fusion peptide or antigen-binding fragment thereof.
The method of any of the preceding claims, wherein the nanoparticles have a mean diameter ranging in size between about 10 nm to about 50 nm.
The method of any of the preceding claims, wherein the nanoparticles have a mean diameter of about 20 nm or about 30 nm.
The method of any of the preceding claims, wherein the nanoparticles are coated with an antibody or fragment thereof that specifically binds human Fc protein, protein A or protein G, or a combination thereof.
The method of any of the preceding claims, wherein the solid support is a bead, resin or agarose.
The method of any of the preceding claims, wherein the nanoparticle comprises or consists of a metal or a polymer.
The method of any of the preceding claims, wherein the solid support is coated with an antibody or fragment thereof that specifically binds human Fc protein, protein A or protein G, or a combination thereof.
The method of any of the preceding claims, wherein the solid support is a bead having a mean diameter at least 2×, 5×, 10×, 50×, 100×, 500×, or 1000× that of the nanoparticle.
The method of any of the preceding claims, wherein the solid support is a bead having a mean diameter ranging in size between about 1 μm to about 10 μm or between about 6.5 μm to about 10 μm.
The method of any of the preceding claims, wherein the fluidic device comprises or consists of a microfluidic chip or sequestration pen.
The method of any of the preceding claims, wherein a single clone of cells are seeded into the fluidic device.
The method of any of the preceding claims, wherein the optical signal comprises light emission, optical pattern, or light scattering.
The method of any of the preceding claims, wherein the change in optical signal is detected using an emission filter.
The method of any of the preceding claims, wherein the change in optical signal is detected using fluorescence scan.
A system comprising:
- a fluidic device;
- nanoparticles coated with a ligand for an antibody protein product;
- and
- solid supports coated with a ligand for the antibody protein product,
- optionally wherein a mean diameter of the solid supports is greater than that of the nanoparticles.
Dependencies are inferred from the claim text.