Method for evaluating blocking activity of immune checkpoint molecular blocking antibody by magnetic beads

Abstract

The invention belongs to the field of immunological detection and particularly relates to a method for evaluating the blocking activity of an immune checkpoint molecular blocking antibody by magneticbeads. Magnetic beads with solid-phase anti-tag molecular antibodies and immune checkpoint molecular protein expressing label molecules (label I) subjected to solid-phase on the surfaces of the magnetic beads are used; the magnetic beads can be specifically bound with a ligand of an immune checkpoint molecule with another label molecule (label II), and then a label II specific binding antibody labeled by a biologic substance (fluorescent or visible light) is used for quantitatively detecting the immune checkpoint molecular protein / ligand molecular protein compound on the surface of the magnetic bead. When the blocking antibody of the immune checkpoint molecule, the magnetic beads with the immune checkpoint molecule protein in the solid phase and the ligand protein coexist, the blocking activity of the antibody is inversely proportional to the final luminous intensity of the magnetic beads, so that quantitative analysis on the blocking activity of the antibody is realized through lightintensity detection. The magnetic bead method is simple to operate, quick in reaction, economical and time-saving.

Description

technical field [0001] The invention belongs to the field of immunological detection, in particular to a method for evaluating the blocking activity of immune checkpoint molecule blocking antibodies by magnetic beads. Background technique [0002] Immune checkpoint molecules are a class of molecules present in the immune system, which interact with their ligands to transmit co-stimulatory or co-inhibitory signals to immune cells to enhance or inhibit immune responses. The binding of immune checkpoint molecules to their ligands has the characteristics of saturation, high affinity, specificity and reversibility. Under physiological conditions, the binding between immune checkpoint molecules and their ligands is not mediated by covalent bonds, but mainly by ionic bonds, hydrogen bonds, van der Waals forces and hydrophobic interactions. The immune response regulation mediated by immune checkpoint molecules plays an important role in maintaining immune homeostasis and inhibiting...

AI Technical Summary

Problems solved by technology

[0003] From the perspective of the effect on the binding efficiency and activity of immune checkpoint molecular proteins and their ligand molecular proteins: the traditional functional ELISA method uses high protein binding activity on the well surface of the plate and multiple epitopes of immune checkpoint molecular recombinant proteins through hydrophobic groups. The physical adsorption between groups produces random binding, which will have the following two adverse effects on the binding between immune checkpoint molecules and their ligand molecules: the first is due to the uncontrollable direction of random binding A part of immune checkpoint molecules The part of the protein surface that can bind to its ligand is unable to bind to its ligand molecule protein due to the combination with the surface of the well plate, which reduces the binding efficiency between the immune checkpoint molecule and its ligand molecule; the second is the surface of the well plate and the immune checkpoint The strong hydrophobic interaction of the molecular recombinant protein will change its spatial conformation to a certain extent, so that the binding activity between the immune checkpoint molecular recombinant protein and its ligand molecular protein will be affected to a certain extent

Images