MULTIPLEXED CHROMATOGRAPHY-IMMUNOASSAY METHOD FOR THE CHARACTERIZATION OF CIRCULATING IMMUNE COMPLEXES

摘要

Thus, herein is reported a method for analyzing/characterizing circulating immune complexes (CICs) formed in vivo comprising a size-exclusion chromatography of a sample obtained from a mammal to which the drug had been administered at least once for determining the weight/size of the immune complexes, optionally a second non-SEC chromatography, and at least one immunoassay, whereby the immune complex is characterized by the correlation of the immune complex size and the immunoassay result/read-out. Also reported herein is the use of a method as reported herein for determining a correlation to altered pharmacokinetics, for determining loss or reduction of efficacy, for determining neutralization of natural counterparts of the drug, for determining immune and hypersensitivity reactions, including serum sickness/type III hypersensitivity reaction/immune complex-mediated disease.

主权项

1. A method for determining a circulating complexed anti-drug antibody comprising a (exogenous) therapeutic polypeptide and an endogenous anti-drug antibody formed in vivo comprising
a) a size-exclusion chromatography of a sample from a mammal to which a drug had been administered at least once for determining the weight/size of the immune complex, b) optionally a second non-SEC chromatography, c) at least one heterogeneous immunoassay for detecting the anti-drug antibody, and d) optionally a mass-spectrometry-based analysis, whereby the immune complex is characterized by the correlation of immune complex size and immunoassay or mass-spectrometry assay read-out/result, whereby the therapeutic polypeptide is a synthetic or not-naturally occurring therapeutic polypeptide.