Antibody Internalization
Antibodies specific to cell surface antigens can induce endocytosis, leading to the internalization of the antibodies along with any molecules conjugated to them. This process, termed Antibody Internalization (ABI), has powerful applications in both research and clinical discovery, including:
● Delivery of highly toxic drugs to specific cancer cells via antibody-drug conjugates (ADCs)
● Removal or degradation of surface receptors from cancer cells (e.g., EGFR)
● Antibody immunotherapies that identify tumor cells for immune cell-mediated killing (e.g., ADCC or ADCP)
AlpVHHs offers a series of VHH (nanobody)-based nano-secondary products specifically designed for evaluating antibody internalization efficiency during the early stage of ADC drug development. This series is particularly suitable for:
● High-throughput screening of antibody internalization efficiency prior to ADC construction
● Rapid functional evaluation and comparison of multiple antibody candidates
● Early-stage ADC drugability assessment in cell-based systems
All products are based on VHH nano-secondary technology, ensuring consistent secondary recognition and reliable assay performance, making them ideal tools for ADC pre-screening workflows.
Before investing in ADC synthesis, it is critical to determine whether candidate antibodies possess sufficient internalization capability. The AlpVHHs Nano-secondary series enables:
● Functional evaluation of antibody internalization
● Rapid comparison of multiple antibody candidates
● High-throughput screening in cell-based systems
● Early-stage ADC drugability assessment
Accelerate your antibody-drug conjugate (ADC) research with AlpVHHs' advanced nanobody-based tools. Our Nano-Secondary ADCs, Nano-DT detection kits, and pH-sensitive probes provide precise, high-throughput internalization detection for both human and mouse antibodies. Optimize your workflow with low-background, high-specificity reagents trusted by leading biotech and pharmaceutical partners worldwide.
| Product Line | Detection Principle | Target Region | Species | Best For |
|---|---|---|---|---|
| Nano-Secondary ADCs | Cytotoxic payload (MMAE/MMAF/DXd/PBD) → cell-killing readout | Fc or CH1 of IgG | Human, Mouse | Functional/cytotoxicity-based internalization screening before ADC construction |
| Nano-DT | Diphtheria toxin fragment → cell-killing readout | IgG (no cross-reactivity to bovine IgG) | Human, Mouse | High-sensitivity screening for low-expression targets; replacement for protein-G-based DT-3C |
| Nano-pH-sensitive Probe | pH-activatable fluorescent dye → imaging/flow readout | Fc, Fab, CH1 (multiple formats) | Human, Mouse | Live-cell imaging, intracellular trafficking, quantitative endosomal signal |
AlpVHHs nanobody internalization reagents are produced under a quality system built for both research and translational use:
ISO 9001 certified quality management system
5-stage quality control: IQC → IPQC → FQC → OQC → QMS
Backed by our own patented technology: "Method for detecting antibody internalization and use of nanobody-toxin drug conjugate thereof"
Recombinant expression (animal-free production after initial discovery), ensuring high lot-to-lot consistency
For Research Use Only (RUO). Not intended for diagnostic or therapeutic use in humans.
Nano-Secondary ADCs and Nano-DT give you a functional, cytotoxicity-based readout — ideal when you want a go/no-go answer before committing to ADC synthesis. The pH-sensitive probe gives you a real-time, quantitative imaging readout — better suited for mechanistic studies of trafficking kinetics rather than a simple pass/fail screen.
No. Unlike traditional DT-3C (which uses a protein G binding domain and can bind serum IgG, causing interference), Nano-DT uses a high-affinity, high-specificity nanobody that does not bind bovine IgG, reducing background interference.
Yes, each product line offers human- and mouse-specific formats — see the SKU table in each section above.
Yes — contact our technical team with your antibody/target details for a feasibility assessment.
Endotoxin testing is not routinely performed on this product line. If your application requires endotoxin-validated reagents, please contact our technical team to discuss available testing options.
We do not currently offer free samples for this product line. Please contact us for a quote based on your required size and volume.
Yes. All products in this series are For Research Use Only (RUO) and are not intended for diagnostic or therapeutic use.
Talk to Our Technical Team , we'll help you match the right detection format to your assay design, species, and throughput needs.
Nano-secondary antibody-Drug conjugates
AlpVHHs offers a new generation of Nano-Secondary antibody-drug conjugates with MMAE, MMAF, DXD or PBD for antibody internalization assays.
The use of Nano-Secondary antibody-drμg conjμgates (Nano-Secondary ADCs) in cell-based cytotoxicity assays is a rapid and cost-effective alternative for pre-screening candidate ADC monoclonal antibodies against tumor cells.
Nano-Secondary ADCs are highly specific and minimally toxic to cells in the absence of primary antibodies, and there is no significant change in primary antibody activity when the conjμgated secondary antibody is present.
Overview:
● Engineered high-affinity VHH nanobody conjugates
● Target Fc or CH1 fragments of human and mouse IgG
● Ready-to-use for flow cytometry, confocal imaging, and live-cell analysis
Functional Cytotoxicity-Based Internalization Screening
● VHH nano-secondary conjugated with potent cytotoxic payloads: MMAE, MMAF, DM1, DXD, Duocarmycin, PBD
● Evaluate internalization-dependent cytotoxicity
● Compare internalization efficiency among multiple antibody candidates
● Support high-throughput screening in tumor cell models
Ideal for rapid pre-selection of internalizing antibodies before formal ADC development.
Technical Advantages:
● High sensitivity with low background
● Compatible with multiple IgG subtypes
● Simplified workflow for high-throughput assays
| Code | Description | Applications | Size |
| 023-101-101 | Anti-Human IgG Fc, AlpSdAbs® VHH(VcMMAE ×4) | Internalization Test | 100ug |
| 023-101-102 | Anti-Human IgG Fc, AlpSdAbs® VHH(VcMMAF ×4) | Internalization Test | 100ug |
| 023-101-103 | Anti-Human IgG Fc, AlpSdAbs® VHH(GGFG-DXD ×4) | Internalization Test | 100ug |
Nano-DT
DT-3C is a commonly used endocytosis detection reagent, but its binding domain is protein G, which will bind to IgG in serum and cause interference. Secondly, the affinity of protein G to different subtypes of mouse and human IgG is different, which will interfere with the result judgment. Nano-DT replaces the protein G binding domain in DT-3C with a high-affinity, high-specificity nanobody that does not bind to bovine IgG.
Nano-DT is an efficient antibody endocytosis detection tool that can be used as a high-throμghput detection tool for pre-drμg evaluation of ADC-related antibodies.
Overview:
● Nanobody-based detection system enhancing traditional DT-3C assays
● Provides rapid and reliable internalization readout
Sensitive Functional Internalization Assessment
● VHH nano-secondary conjugated diphtheria toxin fragments
● High-sensitivity functional readout of antibody internalization
● Suitable for low-expression targets and early-stage ADC feasibility studies
Provides reliable validation of internalization capability during high-throughput screening.
Technical Advantages:
● Improved signal-to-noise ratio
● Supports diverse antibody types and cell lines
● Ready-to-use reagents to reduce experimental complexity
Nano-pH-sensitive probe
Nano-pH-sensitive probe is a new generation of nanobodies (<15KD) coupled to pH-dependent fluorescent probes for endocytosis detection launched by AlpVHHs. pH-sensitive dyes have no fluorescence excitation outside the cell, but emit bright red or green fluorescence in some acidic environments inside the cell (including phagosomes). Unlike traditional labeling methods using amino or thiol-reactive markers, Nano-pH-sensitive probes, which are site-specifically coupled to pH-dependent fluorescent probes by specific nanobodies, are very stable and compatible with BSA and other stable proteins.
Nano-pH-sensitive probe is a fast and economical antibody endocytosis detection reagent.
Overview:
● Fluoresces in acidic endosomal/lysosomal compartments for real-time monitoring
● Available dyes: Deep Red 630, pH-Red 600
Quantitative Evaluation and Real-Time Imaging
● VHH nano-secondary conjugated pH-sensitive fluorescent probes
● Fluoresces in acidic endosomal/lysosomal compartments
● Enables live-cell imaging and flow cytometry analysis
● Quantitative assessment of antibody internalization efficiency
Perfect for mechanistic studies, trafficking analysis, and high-content imaging during antibody screening.
Technical Advantages:
● Track ADC internalization in live cells
● High specificity reduces background
● Compatible with confocal microscopy and high-throughput assays
| Code | Description | Applications | Size |
| 001-101-014 | Anti-Mouse IgG, AlpSdAbs® VHH(Deep Red630) | Internalization Test | 100ug |
| 023-101-014 | Anti-Human IgG, AlpSdAbs® VHH(Deep Red630) | Internalization Test | 100ug |
