Methods for detecting a neutralizing antibody (nab) against an anti-tnf alpha antibody
OwnerThomas Matthews, Kenneth Swartinventors
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Abstract
The present technology comprises methods for detecting a neutralizing antibody (NAb) against a target antibody. In some embodiments, the target antibody comprises an anti-Tumor Necrosis Factor-alpha (TNFα) antibody. In some embodiments, the target antibody comprises SEQ ID NO: 9-11 or 15-17 and SEQ ID NOs: 12-14 or 18-20. The present technology further comprises kits for performing the methods of the present technology.
Claims74
A method for detecting a neutralizing antibody (Nab) against a target antibody in a sample comprising:
- (i) disassociating the NAb from the target antibody by adding an acid wash to the sample;
- (ii) purifying the NAb in the sample;
- (iii) contacting the purified NAb with a detection antibody comprising an anti-TNFα antibody conjugated to a detection label;
- (iv) contacting the purified NAb and the detection antibody with a TNFα-coated substrate;
- and
- (v) detecting the NAb when the detection antibody binds the NAb instead of the TNFα-coated substrate,
- wherein the target antibody is an anti-TNFα chimeric human-murine monoclonal IgG1-kappa antibody comprising SEQ ID NO:
- 9-11 or 15-17 and SEQ ID NOs:
- 12-14 or 18-20.
A method for detecting an NAb against a target antibody in a sample comprising:
- (i) purifying the NAb in the sample;
- (ii) determining whether the NAb is present in the sample by performing a competitive binding assay, wherein the NAb is present if the NAb binds the anti-TNFα antibody with a higher affinity than the affinity of the anti-TNFα antibody to TNFα;
- and
- (iii) detecting the NAb;
- wherein the target antibody comprises an anti-TNFα chimeric human-murine monoclonal IgG1-kappa antibody comprising SEQ ID NO:
- 9-11 or 15-17 and SEQ ID NOs:
- 12-14 or 18-20, and
- wherein a sensitivity of the method is 250 ng/mL or less.
A kit for detecting a NAb against a target antibody in a sample comprising:
- (i) an affinity antibody comprising the target antibody conjugated to a first affinity label;
- (ii) a first solid phase coated with a second affinity label;
- (iii) a detection antibody comprising the target antibody conjugated to a detection label;
- (iv) a substrate;
- and
- (v) TNFα,
- wherein the target antibody is an anti-TNFα chimeric human-murine monoclonal IgG1-kappa antibody comprising SEQ ID NO:
- 9-11 or 15-17 and SEQ ID NOs:
- 12-14 or 18-20.
Dependencies are inferred from the claim text.