By offering efficient and precise Antibody-directed enzyme prodrug therapy (ADEPT) antitumor analytical services, Creative Biolabs provides major technical support for drug development and cancer treatment research. Whether it involves antibody selection, enzymatic reactions, or drug transition, we can deliver optimal experimental design solutions to suit your needs. Our analysis services encompass a large number of areas, including cell viability assessment, drug metabolism profiling, and tumor cell apoptosis detection, enabling you to obtain a full understanding of ADEPT's antitumor efficacy and uncover potential therapeutic mechanisms.

Introduction

In our ADEPT development platform, Creative Biolabs offers a comprehensive analysis of the antitumor efficacy of ADEPT, enabling clients to accurately assess its effects on cancer cells. We can establish standardized tumor cell models that simulate various tumor microenvironments to ensure the reliability and repeatability of experimental results. Moreover, by using specific antibody-directed enzyme prodrug conjugates, our lab can treat tumor cells under in vitro conditions, facilitating the activation of prodrugs targeted to cancer cells. During the experimental analysis, we employ multiple methods to evaluate anti-tumor effects, including cell viability assays, apoptosis analysis, and cell cycle assessments. By comparing the results of the experimental group with the control group, we can quantify the effectiveness of ADEPT in killing tumor cells. Furthermore, we will monitor potential side effects to ensure the safety of the ADEPT therapy.

Fig.1 The ADEPT Method for Tumor Therapy.^1,3

Services

Nowadays, ADEPT has become a promising targeted treatment strategy, offering new avenues for cancer therapy. Creative Biolabs is equipped to provide a variety of ADEPT antitumor analysis services to clients globally, thereby accelerating the development of ADEPT.

• Cell Uptake and Endocytosis Assays

To assess the binding of antibodies to prodrugs and their subsequent cellular uptake efficiency, our laboratory performs cell uptake and endocytosis experiments. These studies aid in determining the distribution of different antibody or prodrug constructs within target cells.

• Enzyme Activity Assays

Creative Biolabs provides activity assays for specific enzymes to evaluate their performance in simulated ADEPT environments. This includes measuring the efficacy of enzyme-catalyzed conversion of prodrugs into active drugs using fluorescent or colorimetric techniques.

• Target Detection Systems

Our research team uses techniques such as Western blotting, immunofluorescence, and flow cytometry to detect the binding capabilities of antibodies to tumor cell targets, ensuring the specificity and efficacy of the ADEPT approach.

• Cell Proliferation Inhibition Assessment

Creative Biolabs evaluates the inhibitory effects of various doses of prodrugs and conjugates on cancer cells using methods such as MTT assays and colony formation tests.

• Apoptosis and Cell Cycle Analysis

We employ flow cytometry to examine the rates of apoptosis and the distribution of the cell cycle, thereby determining the effects of specific drugs on tumor cells.

• Cell Migration and Invasion Experiments

Based on scratch assays and Transwell assays, we assess the impact of ADEPT treatment on the migration and invasion capabilities of tumor cells. These experiments provide crucial data to support the anti-tumor activity.

• Targeting and Selectivity Testing

Our labs evaluate the targeting capabilities and selectivity of ADEPT within tumor tissues using co-culture models or 3D tumor systems, comparing its effects on normal cells versus cancer cells.

• Signal Transduction Pathway Analysis

We can assist clients in analyzing how ADEPT treatment impacts the signal transduction pathways in tumor cells. This analysis may involve qPCR, RNA sequencing, or proteomics to assess changes in the expression of relevant targets.

• Combination Therapy Experiments Design

Our researchers can design and conduct combination therapy experiments to explore the synergistic effects of ADEPT in conjunction with other treatments, such as chemotherapy and immunotherapy, to identify the most effective combination strategies.

• Resistance Mechanism Investigation Evaluation

Creative Biolabs analyzes tumor cells that develop resistance during ADEPT treatment, aiming to uncover the underlying mechanisms of resistance while assessing potential strategies for improving treatment approaches.

• Biomarker Discovery and Validation

Through genomic and proteomic analyses, we can identify biomarkers associated with responses to ADEPT therapy to facilitate patient selection in future clinical applications.

Fig.2 The Antitumor Analysis of NK Cell Functions.^2,3

Case Study:

a. Experimental Design

At Creative Biolabs, our labs have developed a wide collection of in vitro assays to evaluate the efficacy and specificity of ADEPT across various cancer cell types. Our assays include comparisons between untreated cells, cells treated with a single enzyme, and cells subjected to ADEPT therapy.

b. Experimental Process

1. Cell Culture
   - Human tumor cell lines, such as MCF-7 (breast cancer) and A549 (lung cancer), will be cultured in DMEM supplemented with 10% FBS until they reach the log phase of growth.
2. Preparation of Antibodies and Enzymes
   - Highly specific monoclonal antibodies will be selected and conjugated to an appropriate enzyme (such as amidase) using an antibody binding technique, resulting in the formation of an antibody-enzyme complex.
3. ADR (Antibody-Directed Release) Experiment
   - The antibody-enzyme complex will be co-cultured with tumor cells for 4 hours. Afterward, unbound complexes will be washed away using PBS to ensure the specificity of the signal.
4. Prodrug Treatment
   - Following the treatment with the antibody-enzyme complex, prodrugs resembling ketones (e.g., CAP) will be added to the cells in varying concentrations to assess the impact of different doses on cell viability.
5. Cell Proliferation and Viability Analysis
   - Cell viability will be measured using the MTT assay (or CCK-8 assay). The results will be compared to control groups to calculate cell survival rates and proliferation inhibition rates.
6. Flow Cytometry Analysis
   - Flow cytometry will be employed to evaluate cell cycle progression and apoptosis. This assessment will determine ADEPT's effects on cell cycle dynamics and its effectiveness in inducing apoptosis.

c. Data Analysis

- All data will be compiled and analyzed using statistical software to generate survival curves and Dose-Response curves, thereby evaluating the efficacy of ADEPT.

Features

• Unique Targeted Drug Release Mechanism: ADEPT integrates the dual characteristics of antibodies and enzymes, enabling precise pro-drug release through the targeting of tumor cell surface antigens. Our approach enhances the specificity and efficacy of treatment. Our labs use efficient cell culture and co-culture techniques to simulate drug release in a realistic physiological environment.
• Highly Sensitive Detection Techniques: Utilizing state-of-the-art biomolecular detection platforms, we can quantitatively assess the anti-tumor effects of ADEPT therapy at the cellular level. By employing flow cytometry and fluorescence imaging technologies, we continuously monitor tumor cell viability and apoptosis, ensuring the accuracy and reliability of our data.
• Diverse Experimental Models: Our services encompass a wide variety of tumor cell lines and culture models, catering to different experimental requirements. This flexibility allows clients to evaluate the effectiveness of ADEPT therapy across various biological contexts, laying a solid foundation for subsequent clinical research.
• High Adjustability: During the analysis process, we can modify the ratio of the enzyme to the parent drug according to client requirements, optimizing the drug's release and duration of action. This adaptability facilitates the creation of personalized treatment plans.
• High Sensitivity Detection: By using modern biotechnological methods, Creative Biolabs is capable of real-time monitoring of drug activity and anticancer effects at the cellular level, ensuring the scientific validity and accuracy of experimental data. This is crucial for subsequent drug screening and evaluation.
• Multi-Parameter Evaluation: In our ADEPT anticancer analysis, we employ a multi-parameter evaluation technique that considers factors such as intracellular drug uptake, metabolic pathways, and cytotoxicity. This systematic approach not only provides a comprehensive understanding of the mechanisms of action for antibodies and prodrugs but also offers a scientific basis for optimizing treatment strategies.
• Combination of Quantitative and Qualitative Analysis: We have integrated advanced mass spectrometry techniques, enabling precise quantitative and qualitative analysis of metabolites in tumor cells post-ADEPT treatment. The adoption of this technology allows us to monitor the dynamics of drug release and metabolism at the molecular level, thereby informing drug design and improvements.

Creative Biolabs can precisely evaluate the effects of various antibody and enzyme combinations on tumor cells, ensuring the acquisition of highly reliable data. Our team comprises experienced biologists and medicinal chemists with extensive expertise, enabling us to deliver comprehensive analysis and customized services for the characteristics of ADEPT. Today, our ADEPT antitumor analysis service offers a more targeted and personalized approach compared to traditional drug testing methods, ensuring optimal utilization of the specificity of antibodies and the targeting capabilities of enzymes. We sincerely invite you to contact us for more information about our ADEPT antitumor analysis services. Together, let's advance the innovation in ADEPT development.

References

1. Bonifert, Günther, et al. "Recombinant horseradish peroxidase variants for targeted cancer treatment." Cancer medicine 5.6 (2016): 1194-1203.
2. Dean, Isaac, et al. "Rapid functional impairment of natural killer cells following tumor entry limits anti-tumor immunity." Nature Communications 15.1 (2024): 683.
3. Distributed under Open Access license CC BY 4.0, without modification.

For Research Use Only | Not For Clinical Use