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Your Position: Home > Kits > MMAE > ADC-P005

AGLink® ADC Conjugation Kit (MMAE, DAR2&4, 1mg)

For research use only.

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Materials Provided
BoxA(-20°C)
IDComponentsSize
ADC-P005-C0120 x Reaction buffer55ul
ADC-P005-C02Cofactor A55ul
ADC-P005-C03Cofactor B55ul
ADC-P005-C04Substrate A30ul
ADC-P005-C05Substrate B30ul
ADC-P005-C06Enzyme A30ul
ADC-P005-C07Enzyme B30ul
ADC-P005-C08Enzyme C25ul
 
BoxB(4°C)
IDComponentsSize
ADC-P005-1-C01Binding buffer15ml
ADC-P005-1-C02Elution buffer1.5ml
ADC-P005-1-C03Neutralization buffer0.5ml
ADC-P005-1-C04250 mM Tris-HCl buffer (pH7.5)1.5ml
ADC-P005-1-C0510×PBS (pH7.2- 7.4)6ml
ADC-P005-1-C06Desalting column2per
ADC-P005-1-C07Concentrator tube2per
ADC-P005-1-C08ProteinA Resin100ul×2
  • Background
    Antibody-drug conjugates (ADC) is a type of fast growing anticancer drug. Monoclonal antibody is conjugated to the cytotoxic payload via a chemical linker that directed toward a target antigen expressed on the cancer cell surface, reducing systemic exposure and therefore toxicity. AGLink® ADC conjugation kit is designed to conjugate mAb with toxic payload (MMAE) though site-specific enzymatic reaction.
  • Application

    AGLink® ADC Conjugation Kit (MMAE, DAR2&4, 1mg) is designed to conjugate 1mg mAb with toxic payload (MMAE) through site-specific enzymatic reaction.

    It is for research use only.

  • Features
    1. Site-specific conjugation through Fc-glycan with high conjugation efficiency
    2. Uniform DAR value and highly homogeneous ADC products
    3. No need of amino acid sequence engineering
    4. Easy to use: One-pot & One-step
    5. Two options: DAR2 & DAR4 ADC product
  • Storage
    BoxA: The unopened kit is stable for 12 months from the date of manufacture if stored under temperature ≤ -20°C. The opened kit should be stored according to components table. BoxB: The unopened kit is stable for 12 months from the date of manufacture if stored under 4°C. The opened kit should be stored according to components table.
Assay Principles
 MMAE ASSAY PRINCIPLES

Almost all monoclonal antibodies are glycosylated at (or around) Asn-297 of the Fc domain. While the glycans have different isoforms, the types that typically dominate are G0F, G1F&G2F (>90%)[1]. The A&Glink® Antibody Conjugation Kit utilizes the YTConju™ platform, a glycan remodeling strategy developed by Glyco-therapy Biotechnology Co., Ltd. for the construction of site-specific antibody conjugates[2].

The A&Glink® DAR2&4 Antibody Conjugation Kit integrates the procedure for constructing DAR2 conjugates and DAR4 conjugates in one kit, providing flexibility in DAR design.

To construct the DAR4 conjugates, galactose is first added to the terminal GlcNAc catalyzed by β-1,4-galactosyltransferase (GT) to form LacNAc. Then, α-1,3-fucosyltransferase (FT) recognizes LacNAc and transfers the Fuc-Payload to the GlcNAc of the LacNAc to form a conjugate with a theoretical DAR of 4.

 MMAE ASSAY PRINCIPLES

To construct the DAR2 conjugates, glycans are first removed from the core GlcNAc catalyzed by endoglycosidase. Galactose is then added to the GlcNAc catalyzed by β-1,4-galactosyltransferase (GT) to form core LacNAc. Finally, α-1,3-fucosyltransferase (FT) recognizes LacNAc and transfers the Fuc-Payload to the GlcNAc of the LacNAc to form a conjugate with a theoretical DAR of 2.

The DAR4 and DAR2 conjugation procedures share the same FT, GT and GDP-Fuc-Payload. The main difference between these two procedures is the addition of an endoglycosidase.

These procedures are performed in a “one-pot” manner with only one purification step, which is highly convenient and efficient. Antibody conjugates produced with this kit generally exhibit high homogeneity, high stability and high hydrophilicity.

Typical Data Please refer to DS document for the assay protocol.
 MMAE TYPICAL DATA

ADC DAR value assessment by RP-HPLC

DAR (Drug Antibody Ratio) value of ADC product was analyzed by RP-HPLC (QC tested).

Column: TSK gel Butyl-NPR column (4.6 mm × 35 mm, 2.5μm; TOSOH)

Mobile phase A: 20 mM sodium phosphate, 1.5 M ammonium sulfate (pH 6.9)

Mobile phase B: 75% (v/v) 20 mM sodium phosphate, 25% (v/v) isopropanol (pH 6.9)

Flow rate: 0.4 mL/min

Column temperature: 30 °C Detection wavelength:280nm

Loading Amount:10 µg (5-10 µL)

 MMAE TYPICAL DATA

DAR (Drug Antibody Ratio) value of ADC product was analyzed by RP-HPLC (QC tested).

DAR4 =(1*4.43+2*2.28+3*7.1+4*86.19)/100=3.7505

DAR2=(1*0.52+2*99.48)/100=1.9948

 MMAE TYPICAL DATA

ADC DAR value assessment by RP-HPLC

DAR (Drug Antibody Ratio) value of ADC product was analyzed by RP-HPLC (QC tested).

Column: TSK gel Butyl-NPR column (4.6 mm × 35 mm, 2.5μm; TOSOH)

Mobile phase A:20 mM sodium phosphate, 1.5 M ammonium sulfate (pH 6.9)

Mobile phase B:75% (v/v) 20 mM sodium phosphate, 25% (v/v) isopropanol (pH 6.9)

Flow rate: 0.4 mL/min

Column temperature: 30 °C Detection wavelength:280nm

Loading Amount:10 µg (5-10 µL)

 MMAE TYPICAL DATA

DAR (Drug Antibody Ratio) value of ADC product was analyzed by RP-HPLC (QC tested).

DAR4 =(1*4.43+2*2.28+3*7.1+4*86.19)/100=3.7505

DAR2=(1*0.52+2*99.48)/100=1.9948

  • Statement of Rights
    AGLink products are co-developed by ACROBiosystems Co., Ltd. and Glyco-therapy Biotechnology Co., Ltd. based on Glyco-therapy's conjugation platform (YTConju™) and are not authorized for commercial use. The sale and use of AGLink products doesn't represent any forms of license or authorization of the attached technologies for developing any diagnostic, therapeutic or prophylactic products for commercial purposes.
  • Note of application
    AGLink products are intended for research use only by the Buyer or its users and can not be used for any other purposes, including, but not limited to, use for any clinical diagnostic type use, in vitro or in vivo therapeutic use, prophylactic use, any type of consumption or application to human or animals.
  • Clinical and Translational Updates

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