Many unresolved pathologies with unmet medical needs are caused by complex biologies with multiple targets involved in the respective disease pathway. Multispecific antibodies (MsAbs) have emerged to address multi targeting strategies, aiming to achieve improved drug efficacy. More importantly, they enable new functionalities that cannot be achieved with mixtures of the parental antibodies. As the name indicates, MsAbs contain multiple binding sites to different targets and epitopes in one molecule. The molecular complexity of such molecules may pose significant challenges for their development and clinical use. In this talk, I will give an overview of the analytical methods available to characterize such complex molecules, focusing on mispairing analysis and functional characterization.

Many unresolved pathologies with unmet medical needs are caused by complex biologies with multiple targets involved in the respective disease pathway. Multispecific antibodies (MsAbs) have emerged to address multi targeting strategies, aiming to achieve improved drug efficacy. More importantly, they enable new functionalities that cannot be achieved with mixtures of the parental antibodies. As the name indicates, MsAbs contain multiple binding sites to different targets and epitopes in one molecule. The molecular complexity of such molecules may pose significant challenges for their development and clinical use. In this talk, I will give an overview of the analytical methods available to characterize such complex molecules, focusing on mispairing analysis and functional characterization.

Many unresolved pathologies with unmet medical needs are caused by complex biologies with multiple targets involved in the respective disease pathway. Multispecific antibodies (MsAbs) have emerged to address multi targeting strategies, aiming to achieve improved drug efficacy. More importantly, they enable new functionalities that cannot be achieved with mixtures of the parental antibodies. As the name indicates, MsAbs contain multiple binding sites to different targets and epitopes in one molecule. The molecular complexity of such molecules may pose significant challenges for their development and clinical use. In this talk, I will give an overview of the analytical methods available to characterize such complex molecules, focusing on mispairing analysis and functional characterization.

Antibody-drug conjugates (ADCs) represent a promising drug class that relies on monoclonal antibody recognition of specific cancer-associated antigens for targeted delivery of chemotherapeutic agents. These complex molecules are composed of an antibody linked to a biologically active cytotoxic (anticancer) payload or drug. In most cases, ADCs are internalized upon binding to the cognate antigen and the cytotoxic payload is released, causing cell death. The targeted delivery of cytotoxins to tumor cells allows for the maximum efficacy and minimal toxicity. However, despite the enormous promise, a low therapeutic index has plagued ADC development, particularly for treating solid tumors. Nevertheless, 2019 was a very successful year for the ADC community with newly approved molecules as well as positive clinical interims data showcasing the scientific progress within the field. This scientific progress, its impact on ADC design as well as an outlook to combination therapy with ADCs will be discussed here.

Antibody-drug conjugates (ADCs) represent a promising drug class that relies on monoclonal antibody recognition of specific cancer-associated antigens for targeted delivery of chemotherapeutic agents. These complex molecules are composed of an antibody linked to a biologically active cytotoxic (anticancer) payload or drug. In most cases, ADCs are internalized upon binding to the cognate antigen and the cytotoxic payload is released, causing cell death. The targeted delivery of cytotoxins to tumor cells allows for the maximum efficacy and minimal toxicity. However, despite the enormous promise, a low therapeutic index has plagued ADC development, particularly for treating solid tumors. Nevertheless, 2019 was a very successful year for the ADC community with newly approved molecules as well as positive clinical interims data showcasing the scientific progress within the field. This scientific progress, its impact on ADC design as well as an outlook to combination therapy with ADCs will be discussed here.

Antibody-drug conjugates (ADCs) represent a promising drug class that relies on monoclonal antibody recognition of specific cancer-associated antigens for targeted delivery of chemotherapeutic agents. These complex molecules are composed of an antibody linked to a biologically active cytotoxic (anticancer) payload or drug. In most cases, ADCs are internalized upon binding to the cognate antigen and the cytotoxic payload is released, causing cell death. The targeted delivery of cytotoxins to tumor cells allows for the maximum efficacy and minimal toxicity. However, despite the enormous promise, a low therapeutic index has plagued ADC development, particularly for treating solid tumors. Nevertheless, 2019 was a very successful year for the ADC community with newly approved molecules as well as positive clinical interims data showcasing the scientific progress within the field. This scientific progress, its impact on ADC design as well as an outlook to combination therapy with ADCs will be discussed here.

The selective delivery of a therapeutic agent to the target cell is of major importance, particularly for the development of cancer therapeutics. In recent years, antibody-drug conjugates (ADCs) and immunotoxins were developed which combine selectivity of antibodies with toxicity of cell killing compounds. However, immunotoxins suffer from manufacturability problems and for ADCs, the limited hydrophilicity of cytotoxic compounds is a notorious problem, particularly when antibodies are equipped with a high amount of toxic payloads. Here we report our recent development of an intein-splicing strategy for the generation of immunotoxins based on full length antibodies that displayed remarkable potency. [1] In the ADC field, we established dextran as a modular scaffold for various cargoes, conjugated via enzyme-mediated ligation [2] and “click”-chemistry to a tumour-targeting antibody. Dextran, a biocompatible polysaccharide was modified to form a modular scaffold for delivery of payload molecules without compromising solubility and functionality of the chemoenzymatically attached cell-targeting antibody. This approach provides an access to synthetic antibody-polysaccharide hybrid compounds allowing for multivalency of different cargoes, possessing high drug-to-antibody ratios, remarkable hydrophilicity, and high toxicity in vitro. [3,4]

The selective delivery of a therapeutic agent to the target cell is of major importance, particularly for the development of cancer therapeutics. In recent years, antibody-drug conjugates (ADCs) and immunotoxins were developed which combine selectivity of antibodies with toxicity of cell killing compounds. However, immunotoxins suffer from manufacturability problems and for ADCs, the limited hydrophilicity of cytotoxic compounds is a notorious problem, particularly when antibodies are equipped with a high amount of toxic payloads. Here we report our recent development of an intein-splicing strategy for the generation of immunotoxins based on full length antibodies that displayed remarkable potency. [1] In the ADC field, we established dextran as a modular scaffold for various cargoes, conjugated via enzyme-mediated ligation [2] and “click”-chemistry to a tumour-targeting antibody. Dextran, a biocompatible polysaccharide was modified to form a modular scaffold for delivery of payload molecules without compromising solubility and functionality of the chemoenzymatically attached cell-targeting antibody. This approach provides an access to synthetic antibody-polysaccharide hybrid compounds allowing for multivalency of different cargoes, possessing high drug-to-antibody ratios, remarkable hydrophilicity, and high toxicity in vitro. [3,4]

The selective delivery of a therapeutic agent to the target cell is of major importance, particularly for the development of cancer therapeutics. In recent years, antibody-drug conjugates (ADCs) and immunotoxins were developed which combine selectivity of antibodies with toxicity of cell killing compounds. However, immunotoxins suffer from manufacturability problems and for ADCs, the limited hydrophilicity of cytotoxic compounds is a notorious problem, particularly when antibodies are equipped with a high amount of toxic payloads. Here we report our recent development of an intein-splicing strategy for the generation of immunotoxins based on full length antibodies that displayed remarkable potency. [1] In the ADC field, we established dextran as a modular scaffold for various cargoes, conjugated via enzyme-mediated ligation [2] and “click”-chemistry to a tumour-targeting antibody. Dextran, a biocompatible polysaccharide was modified to form a modular scaffold for delivery of payload molecules without compromising solubility and functionality of the chemoenzymatically attached cell-targeting antibody. This approach provides an access to synthetic antibody-polysaccharide hybrid compounds allowing for multivalency of different cargoes, possessing high drug-to-antibody ratios, remarkable hydrophilicity, and high toxicity in vitro. [3,4]

Multiplexed assay systems allow the analysis of hundreds of molecular parameters in a single experiment. Within the last decade multiplexed immunoassay technologies achieved robust analytical performance and enable to analyze a multitude of proteins from a minute amount of sample. An overview of classical and emerging immunoassays established at NMI & SIGNATOPE ranging from sandwich-, mass spectrometry-, bead array-, and digital array-based immunoassays will be presented. The application of these assay formats will be shown on examples from our collaboration with the Critical Path Institute’s Predictive Safety Testing Consortium (PSTC). Protein safety biomarkers have been analyzed in mouse, rat, dog, monkey, and human with the goal to qualify these markers and to reduce drug-induced organ injury. DigiWest, a novel method developed at NMI allows the detection of hundreds of proteins and protein modifications from a minimal amount of sample. Examples that show that the approach can be used to detect the status of cellular signaling cascades in primary tumors and in patient derived tumor organoids will be given. Prediction of therapeutic responses towards respective treatments together with mechanism-of-action (MoA) analyses are areas were this protein profiling approach might prove to be especially useful.

Multiplexed assay systems allow the analysis of hundreds of molecular parameters in a single experiment. Within the last decade multiplexed immunoassay technologies achieved robust analytical performance and enable to analyze a multitude of proteins from a minute amount of sample. An overview of classical and emerging immunoassays established at NMI & SIGNATOPE ranging from sandwich-, mass spectrometry-, bead array-, and digital array-based immunoassays will be presented. The application of these assay formats will be shown on examples from our collaboration with the Critical Path Institute’s Predictive Safety Testing Consortium (PSTC). Protein safety biomarkers have been analyzed in mouse, rat, dog, monkey, and human with the goal to qualify these markers and to reduce drug-induced organ injury. DigiWest, a novel method developed at NMI allows the detection of hundreds of proteins and protein modifications from a minimal amount of sample. Examples that show that the approach can be used to detect the status of cellular signaling cascades in primary tumors and in patient derived tumor organoids will be given. Prediction of therapeutic responses towards respective treatments together with mechanism-of-action (MoA) analyses are areas were this protein profiling approach might prove to be especially useful.

Multiplexed assay systems allow the analysis of hundreds of molecular parameters in a single experiment. Within the last decade multiplexed immunoassay technologies achieved robust analytical performance and enable to analyze a multitude of proteins from a minute amount of sample. An overview of classical and emerging immunoassays established at NMI & SIGNATOPE ranging from sandwich-, mass spectrometry-, bead array-, and digital array-based immunoassays will be presented. The application of these assay formats will be shown on examples from our collaboration with the Critical Path Institute’s Predictive Safety Testing Consortium (PSTC). Protein safety biomarkers have been analyzed in mouse, rat, dog, monkey, and human with the goal to qualify these markers and to reduce drug-induced organ injury. DigiWest, a novel method developed at NMI allows the detection of hundreds of proteins and protein modifications from a minimal amount of sample. Examples that show that the approach can be used to detect the status of cellular signaling cascades in primary tumors and in patient derived tumor organoids will be given. Prediction of therapeutic responses towards respective treatments together with mechanism-of-action (MoA) analyses are areas were this protein profiling approach might prove to be especially useful.