Select Language

ADC Conjugation Kit — Conjugation with Speed, Precision, and Reliability

Antibody-Drug Conjugates (ADCs) are targeted therapeutics structured as "Antibody-Linker-Payload". By precisely conjugating cytotoxic payloads (e.g., MMAE, DXd) to antibodies, ADCs enable targeted delivery of toxins to tumor cells. Conjugation technology critically influences ADC efficacy, safety, and manufacturability.

We have a new ADC Conjugation Kit, delivering a rapid, precise, and robust ADC preparation process. Generate functional ADCs within 3 hours while maintaining antibody activity and high recovery. The resulting conjugates are suitable for direct use in cellular or animal studies to accelerate candidate ADC screening.

Technology & Principle

Based on interchain cysteine conjugation technology, this kit reduces antibody disulfide bonds to generate reactive cysteine residues for non-site-specific maleimide conjugation. This classic strategy enables rapid, controlled ADC preparation—requiring only 3h to obtain ADC conjugates with homogeneous DAR. The resulting conjugates demonstrate significant cytotoxicity, offering a robust tool for ADC screening and mechanistic studies.

Product Features

Fast: Complete ADC prep in <3 hours

Precise: Controlled DAR 4/8 with preserved antibody activity

Efficient: >95% conjugation, >80% recovery

Low Input: Compatible with ≥2 mg/mL antibodies

Simple: 15 min purification via spin column

Protocol Overview

1. Prepare antibody solution for 2 mg/mL in 1×PBS buffer (pH7.2-7.4)

2. Antibody reduction 37°C, 1h

3. Antibody conjugation 23°C, 1h

4. Reaction quenching RT, 0.5h

5. Purification and buffer change

Product List

Components

The ADC Conjugation Kit (MMAE, DAR4, 200μg, for human IgG1) (Catalog No.: ADC-P013) as an example.

Cat.No.	Components ID	Components	Size (1 Kit)	Storage
Box A ADC-P013	C01	Reducing reagent	1 vial	-20℃
	C02	VcMMAE	1 vial	-20℃
	C03	Quencher reagent	1 vial	-20℃
Box B ADC-P013-1	Buffer01	1xPBS (pH7.2-7.4), 11%Trehalose	5 mL	4℃
Box B ADC-P013-1	Desalting column	Desalting Spin column, 0.5mL	1 per	4℃

Validation Data

The following ADC products were prepared using the ADC Conjugation Kit (MMAE, DAR4, 200μg, for human IgG1, Catalog No.: ADC-P013).

Antibody resource: Trastuzumab biosimilar

DAR and Purity Validation by HPLC (HIC + SEC)

Figure 1. The ADC was prepared using the ADC Conjugation Kit (MMAE, DAR4) and analyzed by HIC and SEC-HPLC. The average drug-antibody ratio (DAR) is 4.0±0.5, and the purity of the ADC is greater than 95%.

DAR Validation by Mass Spectrometry

Figure 2. The DAR (3.82) was calculated from the weighted average of the deconvoluted MS peak areas using LC-MS/MS. The results showed the deconvoluted mass spectra of light chains and heavy chains, and the increase in molecular weight caused by the conjugated payload (1316±3 Da). The heterogeneity in N-glycosylation of heavy chain adds to the complexity of the mass spectrum.

Validation of Antigen Binding Activity

Figure 3. Binding affinity of anti-Her2 antibody and anti-Her2 antibody–MMAE conjugate to human Her2 (Cat. No. HE2-H5225) as determined by BLI (Bio-Layer Interferometry). The conjugate exhibits nanomolar affinity (0.57 nM) for human Her2, comparable to the naked antibody (0.86 nM).

In Vitro Cytotoxicity Validation

Figure 4. In vitro cytotoxicity assays: The ADC can bind and internalize in target cells (SK-BR-3) with high expression of Her2 and release MMAE inside the cells to induce a cytotoxic effect (IC₅₀=0.0058 µg/mL). Meanwhile, no cytotoxicity was observed in Her2 receptor-negative cell lines (MDA-MB-231).

Performance Consistency Across Kit Sizes

Figure 5. In vitro cytotoxicity of products prepared by different size of the ADC Conjugation Kit (Cat. No. ADC-P013, ADC-P014). The result shows very high consistency (RSD＜10%).

Stability Validation

Figure 6. The MMAE-ADC was prepared using the ADC Conjugation Kit, which were left at 37℃ for 21 days. The ADCs were analyzed by HIC and SEC-HPLC. The average drug-antibody ratio (DAR) is 4.0±0.5, and the purity of the ADC is greater than 95%.

For technical support, please contact our dedicated product manager

Related Recommendations

Anti-payload Antibodies

AGLink® Site-specific conjugation kit

ADC Internalization Reagent

ADC Total Solution

Resource Sharing

Webinar playback and appointment

‘ADC Potency Analysis and Quality Control Technology Breakthrough’ Webinar Series

ACROBiosystems proudly presents the flagship 'ADC Potency Analysisand Quality Control Technology Breakthrough' live webinar series. This series will systematicallydeconstruct critical aspects ofADC drug development through five coremodules-from potency analysis to quality control technologiesempowering breakthroughs in ADC therapeutics by overcoming keytechnical bottlenecks.

Watch Now

Addressing Challenges in Antibody-Drug Conjugate Development

Here is the brief recap of the webinar:ADCs have the potential to enable 'precision medicine' with a wide market reach.Challenges in ADCs include managing Target Affinity, enhancing payload conjugation, and assessing Payload Delivery in vivo Pharmacokinetics. We can provide high-quality solutions to address these challenges.

Watch Now

Building the Perfect Antibody-based Therapeutic from Selection to Engineering and Manufacturing

In this webinar, discover how advanced techniques such as Al-driven candidate selection optimization of antibody sequences, and precise conjugation methods come together to address some of the current challenges in antibody-based therapeutics.