Antibody Internalization Analysis Services for Brain Tumors

Monoclonal antibodies are now widely used in anti-brain tumor therapy. An essential property of antibodies is the extent, location, and rate of cellular internalization. The action of antibody-coupled drugs (ADCs) is to kill tumor cells by importing cytotoxic substances into cells to cause cytotoxicity. Their specificity and degree of cellular uptake are critical to the efficacy and safety of ADCs. Therefore, understanding the internalization of antibodies within cells and comparing the uptake of different antibodies is a key requirement for antibody selection and optimization for biological drug discovery.

Our antibody internalization analysis services for brain tumors

The ability to internalize antibodies is an important indicator for the early screening of ADCs drugs against brain tumors. Alfa Cytology provides our customers with efficient and stable antibody internalization assay solutions to help you screen ADCs candidate molecules in a rapid, high-throughput manner. By comparing the quantitative efficacy of the fluorescent signal to the concentration of the antibody-drug molecule, we can effectively expand the detection window and provide a high signal-to-noise ratio and stable quantitative efficacy curve.

In addition, our live cell imaging system and associated antibody internalization reagents provide a simple, integrated and quantitative solution for you to directly study the internalization of antibodies within cells. This solution allows easy comparison of multiple antibodies simultaneously. This approach allows antibody internalization analysis to be performed at an early stage and is valuable for optimizing the efficacy, safety, and pharmacokinetics of novel therapeutic antibodies for brain tumors. In addition, the method is also suitable for understanding the underlying mechanisms of endocytosis and cytokinesis.

Internalized antibody screening services

We can provide our clients with high-throughput screening services for internalized antibodies based on filamentous phage and T7λ phage, and construct phage display antibody libraries and peptide libraries. We can provide screening services for internalized antibodies against a variety of receptors, including EGFR, CTLA4, HER2, CD20, CD3, CD59, CD33, etc.

Internalized antibody screening technology flow

• Construct a secondary antibody library against a certain membrane protein receptor.
• Freshly prepared phage antibodies are mixed with negative cells (membrane protein receptor-) and incubated for a certain period to remove non-specific antibodies that can bind to the membrane protein receptor from the antibody library.
• The supernatant of the incubation mixture is then co-incubated with positive cells (membrane protein receptor+) and the cells are washed several times with a complete culture medium.
• Cells are resuspended in the appropriate amount of culture medium and incubated in a cell incubator at the appropriate temperature for some time to allow internalization of the phage antibodies bound to the cell surface.
• Wash the cells with buffer to remove the phage antibodies that are not internalized but still bound to the cells.
• The cells are freeze-thawed and ultrasonically broken, and the internalized phage is digested by trypsin and infected with E. coli for the next round of amplification and screening.

Technical advantages

• The excellent signal-to-background ratio for flow cytometry analysis.
• Stable internalization quantitative efficacy profile validated on multiple targets such as CD20, Her2, EGFR, Claudin 18.2, etc.
• Target cell library with a variety of brain tumor-associated and overexpressed cell lines to choose from.

Alfa Cytology can provide a complete solution for real-time observation and quantitative analysis of antibody internalization from reagents to quantitative analysis. We combine sensitive kinetic fluorescence measurements of internalization with images to meet your visualization and quantitative analysis needs for antibodies to internalization. Please contact us to discuss your project.