Abstract
Background:
The standard safety evaluation of biotherapeutics includes assessment of immunogenicity. Anti-therapeutic antibodies (ATA) can be detected in serum using immunoassays with a bridging format. However, these assays can be subject to interference.
Results:
In the bridging ATA assay for 3A5 TDC, an antibody-drug conjugate that binds to the multimeric extracellular domain of MUC16 (CA125), soluble CA125 in the serum caused false-positive results by binding to the ATA assay reagents. This interaction was blocked by wheat germ agglutinin lectin as it binds to the glycans in CA125; thus, the specificity of the assay improved.
Conclusion:
The assay development and validation results showed that the addition of wheat germ agglutinin eliminates the interference from circulating CA125 without impacting the ability to detect ATA.
MeSH terms
-
Animals
-
Antibodies, Anti-Idiotypic / blood*
-
Antibodies, Anti-Idiotypic / immunology
-
Antibodies, Monoclonal / administration & dosage
-
Antibodies, Monoclonal / immunology
-
Biological Products / immunology*
-
Biological Products / therapeutic use
-
CA-125 Antigen / immunology*
-
CA-125 Antigen / metabolism
-
Drug Carriers / adverse effects
-
Enzyme-Linked Immunosorbent Assay / methods*
-
False Positive Reactions
-
Humans
-
Immunoassay / methods
-
Immunoconjugates / immunology*
-
Immunoconjugates / therapeutic use
-
Macaca fascicularis
-
Membrane Proteins / immunology*
-
Membrane Proteins / metabolism
-
Neoplasms / immunology
-
Neoplasms / therapy
-
Sensitivity and Specificity
-
Solubility
-
Wheat Germ Agglutinins / immunology
-
Wheat Germ Agglutinins / metabolism*
Substances
-
Antibodies, Anti-Idiotypic
-
Antibodies, Monoclonal
-
Biological Products
-
CA-125 Antigen
-
Drug Carriers
-
Immunoconjugates
-
MUC16 protein, human
-
Membrane Proteins
-
Wheat Germ Agglutinins