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F(ab')2 MMAE Conjugation Kit (With VC-PAB Linkage)

  • Fab MMAE conjugate CM11416
  • MC-VC-PAB MMAE chemical structure
Price:
$990.00
SKU:
CM11416
Availability:
In stock, usually ships within 24 hrs
Shipping:
Calculated at checkout
Quantity:


 

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SDS USER MANUAL CM11416
Kit Data 032919

CellMosaic has designed this kit for labeling any F(ab')2 with monomethyl auristatin E (MMAE) using a valine-citruline p-aminobenzylcarbamate (VC-PAB) linker for early stage screening of your antibody. 

This kit includes maleimide-activated VC-PAB-MMAE, which can be coupled directly to a thiol modified F(ab')2 through alkylation in a single step (similar method developed by Seattle Genetics: Sun et al. 2005, Bioconjugate Chem. 16, 1282-1290). The product is purified to remove any unreacted drug. 

Kit is configured to label either one or three F(ab')2. Reaction scales: 2.2 mg

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Drug Information

  • Name: Monomethyl auristatin E (MMAE) with Mal-Val-Cit-PAB linkage
  • CAS number: 646502-53-6
  • Mechanism of action: Inhibits cell division by blocking the polymerization of tubulin. VC-PAB linker is stable in extracellular fluid, but cleaved by cathepsin B once inside the tumor cell, thus activating the antimitotic mechanism
  • Activities: Antioxidant, anti-inflammatory, anticancer, and insecticidal activities

Requirement for F(ab')2

  1. Preferably >90% pure by gel electrophoresis
  2. Total amount: 2.2 mg (protein content measured by UV. If you don't have the exact amount, please contact CellMosaic for customization.)

Key Features of this PerKit™

  • Offers a simple and easy way for site specific labeling of F(ab')2 with MMAE with minimum exposure to the toxin
  • Cathepsin B cleavable VC-PAB (Ref. Doronina et. al. 2008, Bioconjugate Chem. 19, 1960-1963)
  • Fast and easy preparation: 6 h preparation and <2 h hands-on time
  • All reagents and supplies included for preparation and purification
  • Options to choose tailored services at CellMosaic after conjugation

After conjugation: you can choose to send your sample to CellMosaic for HPLC analysis and determination of the loading.

 


Examples

F(ab’)2 information: F(ab’)2 was prepared using CellMosaic’s PerKit™ F(ab’)2 preparation kit (Cat#: CM51408) from a mouse IgG1 antibody.

Kit Lot number: 5525.S13.032919

Scale of the reaction: 0.5 mg

DAR of the final conjugates: 2 calculated based on the UV absorbance ratio (R) of the conjugate at 248 nm and 280 nm.                      

 

Figure 1. SEC HPLC analysis of F(ab')2 and purified F(ab')2-MMAE conjugates (UV at 220 nm)

figure-1.png

 

 Figure 2. HIC HPLC analysis of F(ab')2 and purified F(ab')2-MMAE conjugates (UV at 205 nm)

figure-2.png

 


Related Products

Thiol Assay Kit (Cat. CM90004)The average free thiol groups per antibody is 4 after reducing. If necessary, you can purchase this kit separately from CellMosaic to measure the free thiols. Total assay time is 20 minutes.

PerKit™ F(ab')2 Preparation Kit (Cat. CM51408)

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