Antibody-directed enzyme prodrug therapy (ADEPT) has emerged as a key focus in cancer treatment due to its high specificity and minimal side effects. Creative Biolabs offers precise ADEPT in vitro testing services to clients worldwide, which includes many ADEPT bioactivity assays to support your research and product development efforts in this area.

Introduction

ADEPT combines specific antibodies with enzymes to achieve targeted drug activation at the tumor site. In the context of ADEPT's in vitro experimental services, bioactivity testing plays a key role, particularly involving the assessment of enzyme catalytic activity, lymphocyte proliferation, and cytotoxicity assays. Bioactivity testing can evaluate the interaction capabilities of antibodies and enzymes, ensuring their effective binding at the cellular level for optimal drug release. Additionally, these tests assist research teams in promptly identifying and addressing potential resistance issues, thereby refining the ADEPT strategy.

During the in vitro phase of the ADEPT biological activity testing, Creative Biolabs will engage the selected antibodies with cell models to assess their binding capabilities and cytotoxic effects. Moreover, our laboratory will conduct enzyme activity assays to quantitatively analyze the activity of enzymes involved in the pro-drug conversion process. In our labs, various techniques such as colorimetric assays, fluorescence methods, or mass spectrometry will be employed to monitor the conversion efficiency. Furthermore, we will measure the concentration of active drugs released from the pro-drug and evaluate their biological activity to determine toxicity to target cells.

Fig.1 Mass Spectrometry Systems for Biological Activity Testing.^1,3

Services

Currently, Creative Biolabs offers a full range of ADEPT biological activity testing services, which primarily encompass three core modules: enzyme catalytic activity assessment, lymphocyte proliferation, and cytotoxicity testing.

• Enzyme Catalytic Activity Assessment Service

At our company, our enzyme catalytic activity assessment system employs state-of-the-art detection methods to efficiently and accurately evaluate the catalytic activity directed at specific enzymes. Our enzyme catalytic activity assessment module employs advanced experimental techniques to accurately determine the catalytic efficiency and specificity of enzymes under various conditions. By optimizing experimental design, Creative Biolabs provides highly reproducible and sensitive detection results, allowing you to gain deep insights into the mechanical activity of the enzymes and their interactions with antibodies.

• Lymphocyte Proliferation Service

Our lymphocyte proliferation module uses innovative techniques to sensitively assess the extent of cell proliferation. By comparing various stimulation conditions, we investigate how different molecules influence lymphocyte growth. Creative Biolabs can evaluate the impact of ADEPT therapy on the activity and proliferation capability of immune cells. Utilizing sensitive flow cytometry and fluorescence labeling techniques, we can perform a detailed analysis of lymphocyte proliferation dynamics under different conditions, revealing how the binding of antibodies to enzymes modulates immune responses.

• Cytotoxicity Testing Service

At Creative Biolabs, our cytotoxicity testing module employs multiple cell lines for a thorough assessment of the toxicity of antibody-directed enzyme prodrugs. Our cytotoxicity assay module focuses on assessing the tumoricidal effects of the ADEPT system on cancer cells. By employing various cell viability assessment methods, we can precisely quantify the toxic effects of different drug concentrations on target cells, helping you identify the optimal treatment dosage and timing.

• Apoptosis Assessment Service

Using flow cytometry in conjunction with Annexin V/PI staining, our laboratory conducts quantitative analysis of the apoptotic process in cells. This method allows us to distinguish between early apoptotic cells and late apoptotic or necrotic cells, providing deeper insights into the impact of ADEPT on cellular fate.

• Reactive Oxygen Species (ROS) Detection Service

To evaluate the oxidative stress response induced by ADEPT and its related enzymes within cells, our team employs specific fluorescent probes to detect the generation of reactive oxygen species. This technique enables us to understand how ADEPT influences the redox state of cells and its potential mechanisms of damage.

• Gene/Cell Expression Testing Service

Through immunofluorescence techniques, we can observe changes in the expression of specific markers to assess the impact of ADEPT on cellular signaling pathways. Meanwhile, by PCR quantitatively analyzing the expression levels of specific genes, Creative Biolabs can reveal the role of ADEPT during the transcriptional phase and its underlying mechanisms.

Fig.2 CCK-8 Assays for Cell Viability Analysis.^2,3

Case Study:

A. The objective of the Experiment

The goal of this study is to evaluate the bioactivity of the ADEPT developed by Creative Biolabs under varying concentrations of prodrugs. The focus of the research is to determine the cytotoxic effects of this combination in specific tumor cell lines.

B. Experimental Design

1. Selection of Cell Lines: Choose representative tumor cell lines, such as the human breast cancer cell line MCF-7.
2. Preparation of Antibody-Enzyme Complex: Using a self-developed and optimized purification process to obtain a highly pure antibody-enzyme complex.
3. Prodrug Preparation: Select appropriate prodrugs that can be activated by enzymes within the cell.
4. Bioactivity Testing:
   - Culture MCF-7 cells in a 96-well plate.
   - Introduce different concentrations of the antibody-enzyme complex.
   - Administer the prodrug at various time points to observe the enzyme activation effect.
   - Assess cell viability using the MTT assay, measuring absorbance changes through colorimetric analysis.

C. Data Analysis

The results indicate that as the concentration of the antibody-enzyme complex increases, there is a significant decrease in cell viability. Also, varying concentrations of prodrugs show marked differences in cell death rates. Our data has demonstrated that the antibody-enzyme complex effectively activates the prodrugs, leading to the death of tumor cells.

Features

• Efficient Experimental Design: Our scientists customize experimental plans to meet your research needs, ensuring the reliability and applicability of your results.
• Leading Experimental Techniques: Our labs employ the latest technological methods to guarantee high precision and signal-to-noise ratios in our detection results, enabling you to achieve clear assessments of biological activity.
• Expert Data Analysis: Creative Biolabs offers thorough data processing and interpretation, allowing you to gain a comprehensive understanding of your experimental outcomes and facilitate the progress of your subsequent research.
• One-Stop Service: From experimental design to final reporting, our staff provides comprehensive support services, relieving you of any concerns along the way.

Creative Biolabs guarantees highly accurate testing of ADEPT's biological activity. In our labs, we offer a diverse range of ADEPT bioactivity testing services to assist researchers in evaluating the effectiveness of various enzyme and prodrug combinations. Whether you're looking to validate new drug targets or assess the efficacy of existing ADEPTs, our ADEPT bioactivity testing services are your perfect solution. If you're interested in our ADEPT bioactivity testing services or have any questions, please don't hesitate to contact us at any time.

References

1. Alcázar Magaña, Armando, et al. "Integrating High-Resolution Mass Spectral Data, Bioassays and Computational Models to Annotate Bioactives in Botanical Extracts: Case Study Analysis of C. asiatica Extract Associates Dicaffeoylquinic Acids with Protection against Amyloid-β Toxicity." Molecules 29.4 (2024): 838.
2. Liatsou, Ioanna, et al. "Development and therapeutic evaluation of 5D3 (CC-MLN8237) 3.2 antibody-theranostic conjugates for PSMA-positive prostate cancer therapy." Frontiers in Pharmacology 15 (2024): 1385598.
3. Distributed under Open Access license CC BY 4.0, without modification.

For Research Use Only | Not For Clinical Use