title: Development and characterization of a pre-treatment procedure to eliminate human monoclonal antibody therapeutic drug and matrix interference in cell-based functional neutralizing antibody assays.
abstract: Biological therapeutics can induce an undesirable immune response resulting in the formation of anti-drug antibodies (ADA), including neutralizing antibodies (NAbs). Functional (usually cell-based) NAb assays are preferred to determine NAb presence in patient serum, but are often subject to interferences from numerous serum factors, such as growth factors and disease-related cytokines. Many functional cell-based NAb assays are essentially drug concentration assays that imply the presence of NAbs by the detection of small changes in functional drug concentration. Any drug contained in the test sample will increase the total amount of drug in the assay, thus reducing the sensitivity of NAb detection. Biotin-drug Extraction with Acid Dissociation (BEAD) has been successfully applied to extract ADA, thereby removing drug and other interfering factors from human serum samples. However, to date there has been no report to estimate the residual drug level after BEAD treatment when the drug itself is a human monoclonal antibody; mainly due to the limitation of traditional ligand-binding assays. Here we describe a universal BEAD optimization procedure for human monoclonal antibody (mAb) drugs by using a LC-MS/MS method to simultaneously measure drug (a mutant human IgG4), NAb positive control (a mouse IgG), and endogenous human IgGs as an indicator of nonspecific carry-over in the BEAD eluate. This is the first report demonstrating that residual human mAb drug level in clinical sample can be measured after BEAD pre-treatment, which is critical for further BEAD procedure optimization and downstream immunogenicity testing.
authors.affiliation: Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08540, United States. Electronic address: weifeng.xu@bms.com.
authors.forename: Weifeng
authors.initials: W
authors.lastname: Xu
authors.affiliation: Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08540, United States.
authors.forename: Hao
authors.initials: H
authors.lastname: Jiang
authors.affiliation: Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08540, United States.
authors.forename: Craig
authors.initials: C
authors.lastname: Titsch
authors.affiliation: Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08540, United States.
authors.forename: Jonathan R
authors.initials: JR
authors.lastname: Haulenbeek
authors.affiliation: Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08540, United States.
authors.forename: Renuka C
authors.initials: RC
authors.lastname: Pillutla
authors.affiliation: Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08540, United States.
authors.forename: Anne-Françoise
authors.initials: AF
authors.lastname: Aubry
authors.affiliation: Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08540, United States.
authors.forename: Binodh S
authors.initials: BS
authors.lastname: DeSilva
authors.affiliation: Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08540, United States.
authors.forename: Mark E
authors.initials: ME
authors.lastname: Arnold
authors.affiliation: Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08540, United States.
authors.forename: Jianing
authors.initials: J
authors.lastname: Zeng
authors.affiliation: Analytical and Bioanalytical Development, Bristol-Myers Squibb, Princeton, NJ 08540, United States. Electronic address: robert.dodge@bms.com.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Dodge
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D057134:Antibodies, Neutralizing / Q000276:immunology*; D000917:Antibody Formation / Q000276:immunology; D001681:Biological Assay / Q000379:methods*; D045744:Cell Line, Tumor; D003429:Cross Reactions / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D009500:Neutralization Tests / Q000379:methods; D044967:Serum / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D007074:Immunoglobulin G
keywords: Acid dissociation; Bioassay; LC-MS/MS; Monoclonal antibody therapeutics; Neutralizing antibody; Sample pre-treatment