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| ID | Components | Size |
| ADC-P004-C01 | 20 x Reaction buffer | 70 μL |
| ADC-P004-C02 | Cofactor A | 40 μL |
| ADC-P004-C03 | Cofactor B | 40 μL |
| ADC-P004-C04 | Substrate A | 70 μL |
| ADC-P004-C05 | Substrate B | 60 μL |
| ADC-P004-C06 | Enzyme A | 270 μL |
| ADC-P004-C07 | Enzyme B | 120 μL |
| ADC-P004-C08 | Enzyme C | 40 μL |
| ADC-P004-C09 | Enzyme D | 100 μL |
| ID | Components | Size |
| ADC-P004-1-C01 | Binding buffer | 15 mL |
| ADC-P004-1-C02 | Elution buffer | 2.1 mL |
| ADC-P004-1-C03 | Neutralization buffer | 0.5 mL |
| ADC-P004-1-C04 | 250 mM Tris-HCl buffer (pH7.5) | 1.5 mL |
| ADC-P004-1-C05 | 10×PBS buffer (pH7.2-7.4) | 6 mL |
| ADC-P004-1-C06 | Desalting column | 2 per |
| ADC-P004-1-C07 | Concentrator tube | 4 per |
| ADC-P004-1-C08 | ProteinA Resin | 2 per |
AGLink® ADC Conjugation Kit (MMAE, DAR2&DAR4, 5mg) is designed to conjugate 5mg mAb with toxic payload (MMAE) through site-specific enzymatic reaction. In addition, it has been demonstrated that the kit can also be used in small-volume studies that pair 1mg monoclonal antibody with toxic payload (MMAE).
It is for research use only.
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.
Almost all monoclonal antibodies are glycosylated at Asn-297 of Fc domain. While the glycans have different isoforms, the types which typically dominate are G0F, G1F&G2F (＞90%). AGlink® ADC kit, which is based on the YTConju™ Platform of Glyco-therapy Biotechnology, employs the N-linked glycosylation site for site-specific toxin conjugation. Firstly, galactose is added to the terminal GlcNAc catalyzed by β-1,4-galactosyltransferase (GT) to form LacNAc. Then, the α-1,3-fucosyltransferase (FT) recognize LacNAc, and the toxin is transferred to the GlcNAc of LacNAc. So, the theoretical DAR value of the final conjugates is 2.
The conjugation procedure, which is enzyme catalyzed, is performed as one-pot by combining glycan modification and toxin conjugation simultaneously. The conjugation procedure, which is easy to operate, is performed under physiological conditions, and organic solvents are not introduced. The conjugates prepared through this method usually have high homogeneity and the binding activity of the conjugates to related antigens is not changed.
Almost all monoclonal antibodies are glycosylated at Asn-297 of Fc domain. While the glycans have different isoforms, the types which typically dominate are G0F, G1F&G2F (＞90%). AGlink® ADC kit, which is based on the YTConju™ Platform of Glyco-therapy Biotechnology, employs the N-linked glycosylation site for site-specific toxin conjugation. Firstly, galactose is added to the terminal GlcNAc catalyzed by β-1,4-galactosyltransferase (GT) to form LacNAc. Then, the α-1,3-fucosyltransferase (FT) recognize LacNAc, and the toxin is transferred to the GlcNAc of LacNAc. So, the theoretical DAR value of the final conjugates is 4.
The conjugation procedure, which is enzyme catalyzed, is performed as one-pot by combining glycan modification and toxin conjugation simultaneously. The conjugation procedure, which is easy to operate, is performed under physiological conditions, and organic solvents are not introduced. The conjugates prepared through this method usually have high homogeneity and the binding activity of the conjugates to related antigens is not changed.
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)
DAR (Drug Antibody Ratio) value of ADC product was analyzed by RP-HPLC (QC tested).
DAR = (0*0+0.4*1+95.33*2)/100=1.91
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)
DAR (Drug Antibody Ratio) value of ADC product was analyzed by RP-HPLC (QC tested).
DAR = (3.35*0+3.41*1+4.7*2+6.28*3+82.25*4)/100=3.61
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