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ADC Control Kit with VC-PAB Linker

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Price:
$560.00
SKU:
CM11429
Brand:
Availability:
In stock, usually ships within 1-3 days
Shipping:
Calculated at checkout
Quantity:


 

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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

  • NameMc-VC-PAB-OH
  • CAS number159857-80-4
  • Chemical formula:  C28H40N6O7
  • MW572.7 Da

Requirement for Antibody

  1. Preferably >90% pure by gel electrophoresis
  2. 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

synthetic-route-cm11429.png


 Protocol

scheme-cm11429.png


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). 

hic-analysis-of-example-control-adc-cm11429.png

 

Figure 2. SEC HPLC analysis of IgG1 (blue trace) and purified IgG1 labeled with VC-PAB linker (red trace).

sec-of-example-control-adc-cm11429.png

 

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.

 

Other Details

Reaction Scale:
1-3 mg
Linker Labeling Sites:
Reduced thiols at heavy chains
Linkage:
stable thiol ether bond