Product Description
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| SDS | USER MANUAL |
This kit is designed to label any IgG antibody with VC-PAB linker. The final labeled product can be used as a negative control for any ADC with VC-PAB linker via thiol labeling chemistry. The preparation is essentially the same as standard ADC preparation with VC-PAB linker. The user supplies the antibody. The kit includes maleimide-activated VC-PAB-OH, which can be coupled directly to the antibody after reduction through alkylation in a single step (a method developed by Seattle Genetics: Sun et al. 2005, Bioconjugate Chem. 16, 1282-1290). The product is purified to remove any unreacted linkers.
This kit provides materials to conjugate 1-3 mg of one (CM11429) or three (CM11429x3) antibody samples (IgG).
If you have less than 1 mg of antibody, please add Antibody PerKit™ Customization for Lower Amount (AS1003) along with this kit.
If you would like CellMosaic to perform the conjugation instead, please add PerKit™ Custom Conjugation (AS1002) along with this kit.
Shipping policy: Heat sensitive and ship with cold packs, choose overnight (ships Mon.-Thur.) or 2-day delivery (ships Mon.-Wed.) at checkout.
Linker Information
- Name: Mc-VC-PAB-OH
- CAS number: 159857-80-4
- Chemical formula: C28H40N6O7
- MW: 572.7 Da
Requirement for Antibody
- Preferably >90% pure by gel electrophoresis
- Total amount: 1-3 mg protein content measured by UV
Key Features of this PerKit™
- Offers a simple and easy way to prepare negative control ADC with cathepsin B cleavable VC-PAB linkage (Ref. Doronina et al. 2008, Bioconjugate Chem. 19, 1960-1963)
- Stable linkage
- Fast and easy preparation: 6 h preparation with <2 h hands-on time
- All reagents and supplies included for preparation and purification
- More than 95% labeled products (free of unreacted linker)
- Options to choose tailored services at CellMosaic after conjugation
After conjugation: you can choose to send your sample to CellMosaic for HPLC analysis and calculation of the loading.
Conjugation Chemistry
Protocol
Example of Preparing Control ADC
Antibody information: A therapeutic antibody (human IgG1 subtype)
Kit Lot number: S313.S9.022221
Scale of the reaction: 1.42 mg of antibody (protein content)
Figure 1: HIC HPLC analysis of IgG1 (blue trace) and purified IgG1 labeled with VC-PAB linker (red trace).
Figure 2. SEC HPLC analysis of IgG1 (blue trace) and purified IgG1 labeled with VC-PAB linker (red trace).
Summary of the results:
|
R value (SEC and HIC) |
0.75 |
|
Average LAR based on R value |
3.2 |
|
Average LAR based on HIC data |
3.0 |
|
Extent of antibody aggregation (%) |
0 |
|
Unreacted antibody (%) |
0 |
|
Unreacted Mc-VC-PAB-OH (%) |
0 |
|
Recovery (%) |
82 |
LAR: Linker-to-Antibody Ratio (LAR): check the user manual other consideration regarding the LAR calculation.
