Current Mass Spectrometric Tools for the Bioanalyses of Therapeutic Monoclonal Antibodies and Antibody-Drug Conjugates

Abstract

The increase in the use of therapeutic monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs) has made the detailed bioanalysis of these drugs essential not only for planning optimal therapeutic programs for clinical practice, but also for evaluating the biological equivalencies in the development of other biosimilars. The ligand binding assays that are widely in use now are being replaced rapidly by the highly accurate, sensitive, and selective analytical method using a mass spectrometer. This review will discuss the progress in and challenges observed during the development of a mass spectrometry-based bioanalytical method for therapeutic mAbs and ADCs.

Analytical Comparison of Antibody-drug Conjugates Based on Good Manufacturing Practice Strategies

“…[1][2][3] In accordance with the growth of the ADC field, analytical characterization continues to play a crucial role in the discovery, development, and manufacturing of ADCs. 4,5 The FDA in particular, requires the pharmaceutical industry to carefully analyze antibody-based biopharmaceuticals, 6,7 which remains a challenge due to the heterogeneity and structural complexity of the compounds. Several analytical methods have been established in an effort to address this issue.…”

“…Several analytical methods have been established in an effort to address this issue. 4,5 High performance liquid chromatography (HPLC) analysis is the most popular analytical approach for drug antibody ratio (DAR) characterization, but it is important to develop multiple methods for application at different project stages to ensure accuracy of DAR determination. Previously, we reported analytical comparison for site-specific ADCs using six analytical techniques to establish the appropriate analytical method for each phase in the project cycle.…”

High-Temperature Reversed-Phase LC Separation of Heavy and Light Chain Fragments of Therapeutic Monoclonal Antibodies and Antibody-Drug Conjugate Produced by Chemical Reduction

“…With the increasing use of therapeutic monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs), there is an increasing demand for the bioanalysis of these drugs, in order to evaluate their efficacies and establish optimal treatment plans. For the bioanalysis of therapeutic mAbs and ADCs in complex biological samples such as plasma and serum, tryptic digestion-liquid chromatography-tandem mass spectrometry (LC-MS-MS) and ligand-binding assays such as enzyme-linked immunosorbent assay are the two most commonly used strategies, primarily because of their high sensitivities and specificities [1,2]. These methods require complex and time-consuming pretreatment, precision control, and homogeneity of the reagents used [2].…”

“…For the bioanalysis of therapeutic mAbs and ADCs in complex biological samples such as plasma and serum, tryptic digestion-liquid chromatography-tandem mass spectrometry (LC-MS-MS) and ligand-binding assays such as enzyme-linked immunosorbent assay are the two most commonly used strategies, primarily because of their high sensitivities and specificities [1,2]. These methods require complex and time-consuming pretreatment, precision control, and homogeneity of the reagents used [2]. Therefore, a new trend has emerged for the direct analysis of the target drugs or their light chain fragments by LC, in conjugation with high-resolution mass spectrometry [3,4].…”

Biosensors Using an Antibody as a Recognition Element

“…The development of a mass spectrometry-based bioanalytical method for therapeutic monoclonal antibodies and antibody-drug conjugates has also been reviewed. 15 The development of novel signal conversion systems utilizing an antibody with excellent properties is of great interest because it permits the simple, rapid, and sensitive sensing of proteins related to diseases, environmental pollutants and pathogens.…”