Immunogenicity Considerations for Antibody–drug Conjugates: A Focus on Neutralizing Antibody Assays

Fiorotti, Corinna

doi:10.4155/bio-2017-0229

Cited by 7 publications

(4 citation statements)

References 35 publications

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“…In general, ADAs elicited for an ADC includes those specific to intact ADC, to linker, to payload, and to the unconjugated antibody. All the ADAs generated against the ADC should be properly detected [ 43 ]. Usually, the entire ADC molecule is used in an ADA assay, enabling identification of ADAs with specificity for any component of the intact ADC.…”

Section: Detection Of Adasmentioning

confidence: 99%

“…Presently, bridging ligand binding assay (Bridging LBA) is the format most commonly used in immunogenicity testing [ 15 , 16 , 43 , 44 ] where a biotinylated drug is utilized for capturing ADAs, and ruthenium or HRP-labeled drug and even digoxigenin (DIG)-labeled drug for detecting the bridged ADA–drug complex (see Figure 3 A). Other assay formats used in immunogenicity testing include direct and indirect assay formats.…”

Section: Detection Of Adasmentioning

confidence: 99%

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Current Analytical Strategies for Antibody–Drug Conjugates in Biomatrices

Qin

Gong

2022

Molecules

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Antibody–drug conjugates (ADCs) are a new class of biotherapeutics, consisting of a cytotoxic payload covalently bound to an antibody by a linker. Ligand-binding assay (LBA) and liquid chromatography-mass spectrometry (LC-MS) are the favored techniques for the analysis of ADCs in biomatrices. The goal of our review is to provide current strategies related to a series of bioanalytical assays for pharmacokinetics (PK) and anti-drug antibody (ADA) assessments. Furthermore, the strengths and limitations of LBA and LC-MS platforms are compared. Finally, potential factors that affect the performance of the developed assays are also provided. It is hoped that the review can provide valuable insights to bioanalytical scientists on the use of an integrated analytical strategy involving LBA and LC–MS for the bioanalysis of ADCs and related immunogenicity evaluation.

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Section: Detection Of Adasmentioning

confidence: 99%

Section: Detection Of Adasmentioning

confidence: 99%

Current Analytical Strategies for Antibody–Drug Conjugates in Biomatrices

Qin

Gong

2022

Molecules

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“…In preclinical studies, ADAs may affect the determination of plasma or serum drug concentrations of ADCs and, thus, impact their pharmacokinetic or toxicokinetic profiles. In clinical practice, ADAs may cause a decrease in or even a loss of efficacy, increased drug toxicity, allergic and hypersensitivity reactions, eventually leading to poor clinical outcomes [ 6 , 9 ]. Therefore, it is suggested that the preclinical ADA results could be used to predict the immunogenicity of drugs in clinical practice.…”

Section: Introductionmentioning

confidence: 99%

Development, Validation and Application of a Bridging ELISA for Detection of Antibodies against GQ1001 in Cynomolgus Monkey Serum

Liu

Sun²,

Deng

et al. 2023

Molecules

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Immunogenicity is a major issue associated with the PK, efficacy, and safety evaluation of therapeutic protein products during pre-clinical and clinical studies. A multi-tiered approach consisting of screening, confirmatory, and titration assays has been widely adopted for anti-drug antibody testing. GQ1001, a recombinant humanized anti-human epidermal growth factor receptor 2 monoclonal antibody covalently linked to a cytotoxin of DM1, possesses a novel format of antibody–drug conjugates. In this study, we reported the development, validation, and application of an acid-dissociation bridging enzyme-linked immunosorbent assay for the detection of antibodies against GQ1001 in cynomolgus monkey serum. The sensitivity of the screening assay was 126.141 ng/mL in undiluted serum. The screening assay and confirmatory assay were neither affected by the naïve monkey serum nor by 2% and 5% (v/v) erythrocyte hemolysates. Moreover, the assay was not subject to interference by 2500 ng/mL of human IgG1 in the samples. Drug interference at low positive control (150 ng/mL) and high positive control (8000 ng/mL) of anti-GQ1001 antibodies was not observed when GQ1001 concentrations were below 3.125 μg/mL and 100 μg/mL, respectively. Furthermore, no hook effect was observed for the positive antibodies in the concentration range of 8 to 64 μg/mL. The validated assay was, thereafter, successfully applied to a single-dose toxicity study of GQ1001. Anti-drug antibody positive rates among dosing animals and testing samples were reported, and no significant impact was found on toxicokinetic outcomes.

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“…In addition, LC–MS/MS assays for small molecules need to be developed to measure unconjugated drugs. Moreover, ADCs are known to elicit anti-drug antibody (ADA) responses, similar to other biologics [ 26 , 27 , 28 , 29 , 30 ]. The ADAs are generated against all components of the intact ADC, which consists of a linker, payload, and unconjugated antibody.…”

Section: Introductionmentioning

confidence: 99%

Implementation of Systematic Bioanalysis of Antibody–Drug Conjugates for Preclinical Pharmacokinetic Study of Ado-Trastuzumab Emtansine (T-DM1) in Rats

et al. 2023

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Antibody–drug conjugates (ADCs) are composed of monoclonal antibodies covalently bound to cytotoxic drugs by a linker. They are designed to selectively bind target antigens and present a promising cancer treatment without the debilitating side effects of conventional chemotherapies. Ado-trastuzumab emtansine (T-DM1) is an ADC that received US FDA approval for the treatment of HER2-positive breast cancer. The purpose of this study was to optimize methods for the quantification of T-DM1 in rats. We optimized four analytical methods: (1) an enzyme-linked immunosorbent assay (ELISA) to quantify the total trastuzumab levels in all drug-to-antibody ratios (DARs), including DAR 0; (2) an ELISA to quantify the conjugated trastuzumab levels in all DARs except DAR 0; (3) an LC–MS/MS analysis to quantify the levels of released DM1; and (4) a bridging ELISA to quantify the level of anti-drug antibodies (ADAs) of T-DM1. We analyzed serum and plasma samples from rats injected intravenously with T-DM1 (20 mg/kg, single dose) using these optimized methods. Based on these applied analytical methods, we evaluated the quantification, pharmacokinetics, and immunogenicity of T-DM1. This study establishes the systematic bioanalysis of ADCs with validated assays, including drug stability in matrix and ADA assay, for future investigation on the efficacy and safety of ADC development.

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Immunogenicity Considerations for Antibody–drug Conjugates: A Focus on Neutralizing Antibody Assays

Cited by 7 publications

References 35 publications

Current Analytical Strategies for Antibody–Drug Conjugates in Biomatrices

Current Analytical Strategies for Antibody–Drug Conjugates in Biomatrices

Development, Validation and Application of a Bridging ELISA for Detection of Antibodies against GQ1001 in Cynomolgus Monkey Serum

Implementation of Systematic Bioanalysis of Antibody–Drug Conjugates for Preclinical Pharmacokinetic Study of Ado-Trastuzumab Emtansine (T-DM1) in Rats

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