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组分（Materials Provided）
BoxA(-20°C)IDComponentsSizeADC-P018-C01Reducing reagent1 vialADC-P018-C02Exatecan1 vialADC-P018-C03Quencher reagent1 vialBoxB(4°C)IDComponentsSizeADC-P018-1-C011xPBS (pH7.2-7.4), 11%Trehalose15 mLADC-P018-1-C02Desalting Spin column 5mL1 per产品概述（Product Overview）
Antibody-drug conjugates (ADCs) deliver potent cytotoxic drugs specifically to tumor cells, thereby enhancing antitumor efficacy and reducing the adverse systemic effects of the cytotoxic drugs. This kit is developed based on interchain cysteine conjugation technology: the interchain disulfide bonds of antibodies can be reduced to generate endogenous interchain cysteine residues, and payload drugs with maleimide functional groups can specifically conjugate to these cysteine residues to form structurally stable ADCs. Relying on this classic conjugation strategy, the kit enables a rapid and controllable ADC preparation process that takes only 3 hours in total; the prepared ADC products have an average drug-to-antibody ratio (DAR) of 8.0±0.5, and the ADC conjugates exhibit significant cytotoxic activity, providing a reliable experimental tool for ADC drug screening and mechanism research.
应用说明（Application）
The ADC Conjugation Kit is designed to conjugate human IgG1 and cytotoxic payloads Exatecan through thiol and maleimide reaction chemistry. The ADC products can be used in cellular or animal experiments for the screening of suitable ADC candidates.
It is for research use only.
产品特性（Features）
- Short Conjugation Time: Generates antibody-drug conjugates (ADCs) within 3 hours.
- Controllable DAR Value: Precisely regulates DAR while well preserving antibody activity.
- High Conjugation Efficiency & Recovery: Conjugation efficiency > 95%, antibody recovery rate > 80%.
- No Need for High-Concentration Antibodies: Reaction feasible with only 2 mg/mL of antibodies.
- Simple Purification: Purification completed in 15 minutes using spin desalting columns.
存储（Storage）
BoxA: This product remains stable when stored at -20℃ for 24 months.
BoxB: This product remains stable when stored at 2-8℃ for 24 months.
Please ensure use prior to the expiration date indicated on the packaging.
质量管理控制体系（QMS）
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数据展示
SEC-HPLC
The purity of ADC product prepared using the ADC Conjugation Kit was greater than 90% as determined by SEC-HPLC.
HIC-HPLC
The Exatecan-ADC was prepared using the ADC Conjugation Kit and analyzed by HIC-HPLC. The average drug-antibody ratio (DAR) is 8.0±0.5.
质谱（Mass Spectrometry）
The DAR (7.92) 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 coupling payload (1149±2 Da). The heterogeneity in N-glycosylation of heavy chain adds to the complexity of the mass spectrum.
活性（Bioactivity）-BLI
Binding affinity of anti-Her2 antibody and anti-Her2 antibody–Exatecan conjugate to human Her2 (Cat. No. HE2-H5225) as determined by BLI (Bio-Layer Interferometry). The conjugate exhibits nanomolar affinity (0.44 nM) for human Her2, comparable to the naked antibody (0.86 nM).
In Vitro Cytotoxic Activity
The ADC can bind and internalize in target cells (SK-BR-3) with high expression of HRE2 and release Exatecan inside the cells to induce a cytotoxic effect (IC50=0.0066 µg/mL). Meanwhile, no cytotoxicity was observed in HER2 receptor-nagetive cell lines (MDA-MB-231).
In vitro cytotoxicity of products prepared by different size of the ADC Conjugation Kit (Cat. No. ADC-P017, ADC-P018). The result shows very high consistency (RSD＜10%).
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