Methods and Systems for Detecting Antibodies by Surface Plasmon Resonance

Patent metadata

Field	Value
Publication number	US-2020333331-A1
Application number	US-202016850541-A
Country	US
Kind code	A1
Filing date	Apr 16, 2020
Priority date	Apr 16, 2019
Publication date	Oct 22, 2020
Grant date	—

Abstract

The present disclosure relates to a method for detection of an antibody or antibody fragment in a biological sample from a subject. In some embodiments, the methods comprise immobilizing a first binding agent on a surface plasmon resonance (SPR) biosensor; adding a ligand that binds to the first binding agent under conditions such that a complex of the ligand and the first binding agent is formed; adding an aliquot of the biological sample under conditions such that the antibody and/or antibody fragment binds to the ligand that is complexed to the first binding agent; and detecting the presence of the antibody and/or antibody fragment as a change in signal obtained from SPR. In some embodiments, the antibody is an antibody therapeutic such as certolizumab pegol. Also disclosed are systems and kits for detecting an antibody in a biological sample from a subject using SPR.

First claim

That which is claimed is:

1 . A method for detection of an antibody or antibody fragment in a biological sample from a subject comprising:
immobilizing a first binding agent on a surface plasmon resonance (SPR) biosensor; adding a ligand that binds to the first binding agent under conditions such that a complex of the ligand and the first binding agent is formed; adding an aliquot of the biological sample under conditions such that the antibody or antibody fragment binds to the ligand that is complexed to the first binding agent; and detecting the presence of the antibody and/or antibody fragment as a change in signal obtained from SPR.

2 . The method of claim 1 , further comprising adding a secondary antibody or a second binding agent that recognizes a moiety on the antibody or antibody fragment prior to the step of detecting the final signal obtained from SPR.

3 . The method of claim 1 , wherein the antibody or antibody fragment is an antibody therapeutic.

4 . The method of claim 3 , wherein the antibody therapeutic is certolizumab pegol.

5 . The method of claim 2 , wherein the moiety on the antibody or antibody fragment is polyethylene glycol (PEG).

6 . The method of claim 1 , wherein the ligand is TNFα.

7 . The method of claim 1 , wherein the first binding agent is an antibody.

8 . The method of claim 1 , wherein the surface plasmon resonance (SPR) provides for real-time detection of the antibody.

9 . The method of claim 1 , wherein the SPR biosensor comprises a sensor chip.

10 . The method of claim 1 , wherein the SPR biosensor comprises a metal surface.

11 . The method of claim 1 , wherein the SPR biosensor comprises a thin alginate layer for amine coupling of ligands.

12 . The method of claim 9 , wherein the sensor chip has a surface coating that can be activated to specifically react with free surface amines of proteins.

13 . The method of claim 1 , wherein the ligand is immobilized at a density of at least 1,000 Response Units (RU).

14 . The method of claim 1 , wherein the biological sample is serum or plasma.

15 . The method of claim 1 , wherein the biological sample is from a human.

16 . The method of any of claim 3 , wherein the biological sample is from a human being treated with the antibody therapeutic.

17 . The method of claim 16 , further comprising adjusting the amount of the antibody therapeutic in the subject based on the amount detected in the biological sample.

18 . The method of claim 1 , wherein the biological sample is diluted at least 2-fold.

19 . The method of claim 4 , wherein the lower limit of quantitation (LLOQ) and upper limit of quantitation (ULOQ) for certolizumab are 1.0 μg/mL and 90 μg/mL, respectively.

20 . A system for detection of an antibody or an antibody fragment in a biological sample from a subject comprising:
a biosensor for surface plasmon resonance (SPR) having a first binding agent immobilized on one surface, wherein the first binding agent is capable of binding to a ligand; the ligand for the first binding agent, wherein the ligand is capable of binding to the antibody or antibody fragment; and a component for measuring a signal from the biosensor for surface plasmon resonance.

21 . The system of claim 20 , further comprising a second antibody or a second binding agent capable of binding to the antibody or antibody fragment.

22 . A kit for detection of an antibody in a biological sample from a subject comprising:
a biosensor for surface plasmon resonance (SPR) having a first binding agent immobilized on one surface; a ligand for the first binding agent, wherein the ligand also binds to the antibody of interest; optionally, a second antibody or a second binding agent that recognizes a moiety on the antibody; and optionally, reagents for washing the biosensor to remove biomolecules that do not specifically bind to the first binding agent, the ligand, or the antibody; and instructions for use.

Assignees

• Laboratory Corp America Holdings

Inventors

• Kelemen Mary Katherine Morr
• Holmquist Brett

Classifications

• G01N21/553

  and using surface plasmons (fluorescence excitation G01N21/648; enhanced Raman G01N21/658) · CPC title

• G01N33/54353

  with ligand attached to the carrier via a chemical coupling agent (coatings G01N33/54393) · CPC title

• G01N33/543 Primary

  with an insoluble carrier for immobilising immunochemicals · CPC title

• G01N33/553

  Metal or metal coated · CPC title

• G01N33/54373

  involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings · CPC title