ADC drugs can not only specifically recognize tumor surface antigens through monoclonal antibodies, but also kill tumor target cells by using high-efficiency small-molecule drug toxins carried by themselves. Combining the advantages of targeted drugs and chemotherapeutic drugs, ADC drugs have gradually become a research hotspot in the past 20 years. However, due to the complex structure, the difficulty of ADC development and production process is greatly increased. Compared with macromolecular drugs, ADC has more diverse degradation modes and toxicity due to the presence of small molecules and linkers, which makes the development, manufacture and biological analysis of ADC face great challenges. The new ADC analysis methodology introduced by BOC Sciences has a wide range of application and diversity, which is crucial for characterizing individual components of ADCs linker, antibodies and payloads. We have developed a series of in vitro and in vivo preclinical assays to help our customers determine the biochemical and in vitro efficacy of ADC and provide comprehensive guidance for project progress.
ADC In Vitro Analysis Abilities
In order to explore the functional effects and mechanism of ADCs, the analysis and verification platform of BOC Sciences comprehensively covers the in vitro evaluation process of ADCs. We can accurately analyze and strictly control the physicochemical properties, pharmaceutical properties, stability and safety of ADC drugs. Our ADC in vitro analysis services include:
- Advanced structural analysis: study the effect of drug conjugation on antibody conformation.
- ADC stability analysis: study the effect of drug conjugation on antibody stability.
- ADC solubility analysis: study the effect of drug conjugation on the solubility of antibody and ADC conjugates.
- Total antibody analysis: bound and unbound antibody analysis - ELISA and electrochemiluminescence immunoassay (ECLA).
- Conjugated antibody ADC analysis: use ELISA and ECLA methods to determine antibody conjugates in serum and perform PK analysis.
- Analysis of small molecule cytotoxicity: orepare standard samples of free small molecule drugs and related metabolites in plasma and synthesize stable isotope-labeled internal standards or simulated molecules, and then use LC–MS/MS analysis method to analyze small molecule breakdown products Nonclinical and clinical studies were conducted.
- Immunogenicity analysis: the standard anti-drug antibodies (ADAs) analysis process was used to detect the immunogenicity of ADC, including screening, confirmation, titer, and neutralization experiments. ADCs have the potential to induce immune responses and produce corresponding ADAs, which in turn affect PK, PD and safety.
- Drug-antibody ratio (DAR) determination: DAR is a critical quality parameter for ADCs, and its analytical characterization techniques play an important role in the discovery, development and manufacture of ADCs. We are equipped with a variety of techniques that can be used to analyze and characterize DAR, including spectroscopic, radioactive, chromatographic, mass spectrometric and more.
In Vitro Efficacy Assessment
- Cytotoxicity assay: cytotoxicity assay is an important method to screen promising ADC candidate cells, predict ADC efficacy in vivo, and assess cell specificity. The effect of cytotoxicity on cell proliferation or killing was determined by MTT method.
- ADC affinity assessment: binding affinity of ADCs to indicated target antigens and antibodies was assessed using surface plasmon resonance (SPR), enzyme-linked immunosorbent assay (ELISA), and fluorescence-activated cell sorting (FACS) flow cytometry.
- Internalization analysis: we have developed a variety of ADC internalization assays to meet different screening needs, including immunotoxin-conjugated killing assays, real-time live cell imaging-based internalization assays, cell surface secondary antibody-based internalization assays, and pH-based probe-tracked internalization assays.
- Antibody dependent cellular cytotoxicity (ADCC) assay: cytotoxicity and cell activity detection methods are used for ADCC in vitro detection, including the determination of target cells (highly expressed monoclonal antibodies on the cell surface can recognize antigens), the selection of effector cells and the detection of target cell activity.
- Complement dependent cytotoxicity (CDC) assay: fluorescent labeling and flow cytometry are used to detect the viability of target cells to evaluate the CDC effect of the drug.
- ADC Targeting: in vitro analysis of ADC targeting capabilities is provided by using cell culture models.
ADC Detection Equipments and Facilities
In order to better control the quality, stability and safety of ADC drugs, BOC Sciences is equipped with a variety of instruments and equipment for ADC drug analysis, which can provide guidance for the analysis, characterization and development of ADC drugs.
- Ultraviolet-Visible Spectroscopy (UV/VIS)
- Mass Spectrometry (MS)
- Hydrophobic Interaction Chromatography (HIC)
- Size Exclusion Chromatography (SEC)
- Isoelectric Focusing (IEF)
- Time-of-Flight (TOF) Analyzer
- Ion Exchange Chromatography (IEC)
- Enzyme-linked immunosorbent assay (ELISA)
- Capillary Electrophoresis (CE)
- High performance liquid chromatography (HPLC)
- Offline Solid Phase Extraction (SPE)
Our ADC Analysis Workflow
BOC Sciences is committed to improving analytical techniques and pairing with appropriate bioanalysis methods, such as ELISA and cell-based potency assays, to help ADC developers better understand how ADC structure affects clinical outcomes such as ADC efficacy and safety. We can help customers choose more suitable ADC candidates to enter clinical research, and provide routine GMP testing and stability monitoring in the mass production of clinical samples. Our cGMP manufacturing capabilities cover cytotoxin and linker synthesis, antibody modification and conjugation, and finished ADC purification and filling.
* Only for research. Not suitable for any diagnostic or therapeutic use.