Tools for ADC PK Analysis——Anti-payload antibodies

Immobilized Mouse Anti-DXD&Exatecan Antibody, Mouse IgG1, mAb (Cat. No. DXD-M684) at 1 μg/mL, add Trastuzum*b deruxtecan in the 50% Human serum and then add Biotinylated Human Her2, His,Avitag, premium grade (Cat. No. HE2-H82E2) at 0.5 μg/mL. Detection was performed using HRP-conjugated Streptavidin (Acro, Cat. No. STN-NH913) (Routinely tested).

Immobilized Human Her2, His Tag (Cat. No. HE2-H5225) at 1 μg/mL, add Trastuzum*b Deruxtecan in the 100% Human Serum and then add Biotinylated Anti-DXD Antibody, Mouse IgG1, Avitag (Cat. No. DXD-BVM807) at 0.05 μg/mL. Detection was performed using HRP-conjugated Streptavidin (Acro, Cat. No. STN-NH913) (QC tested).

Immobilized Mouse Anti-MMAE&MMAF Antibody, Mouse IgG1 (Cat. No. MME-M5252) at 5 μg/mL, add Disitamab Vedotin (RC48) in the 50% Mouse serum and then add Biotinylated Human Her2, His,Avitag, premium grade (Cat. No. HE2-H82E2) at 0.5 μg/mL. Detection was performed using HRP-conjugated Streptavidin (Acro, Cat. No. STN-NH913) (Routinely tested).

Immobilized Monoclonal Anti-DM-1&DM-4 Antibody, Mouse IgG1 (Cat. No. DM1-Y73) at 5 μg/mL, add Trastuzum*b-DM1 in the 50% Human serum and then add Biotinylated Human Her2, His,Avitag, premium grade (Cat. No. HE2-H82E2) at 0.5 μg/mL. Detection was performed using HRP-conjugated Streptavidin (Acro, Cat. No. STN-NH913) (Routinely tested).

The purity of Monoclonal Anti-DXD&Exatecan Antibody, Mouse IgG1 (Cat. No. DXD-M684) is more than 90% and the molecular weight of this protein is around 135-160 kDa verified by SEC-MALS.

The purity of Monoclonal Anti-MMAE&MMAF Antibody, Mouse IgG1 (Cat. No. MME-M5252) is more than 90% and the molecular weight of this protein is around 134-163 kDa verified by SEC-MALS.

Immobilized ADC-DXD at 1 μg/mL (100 μL/well) can bind Mouse Anti-DXD&Exatecan Antibody, Mouse IgG1, mAb (Cat. No. DXD-M684) with a linear range of 0.1-4 ng/mL (Routinely tested).

Immobilized Disitamab Vedotin (RC48) at 0.2 μg/mL (100 μL/well) can bind Monoclonal Anti-MMAE&MMAF Antibody, Mouse IgG1 (Cat. No. MME-M5252) with a linear range of 0.1-2 ng/mL (QC tested).

Immobilized ADC-DM4 at 2 μg/mL (100 μL/well) can bind Monoclonal Anti-DM-1&DM-4 Antibody, Mouse IgG1 (Cat. No. DM1-Y73) with a linear range of 0.20-6.25ng/mL (Routinely tested).

Immobilized ADC-DXD at 1 μg/mL (100 μL/well) can bind HRP conjugated Mouse Anti-DXD&Exatecan Antibody, Mouse IgG1 (Cat. No. DXD-PLM684) with a linear range of 0.5-31 ng/mL (Routinely tested).

ELISA binding of Monoclonal Anti-MMAE&MMAF Antibody, Mouse IgG1 (Cat. No. MME-M5252) with Trastuzumab Deruxtecan, IgG1-MMAF, Sacituzumab Govitecam, Disitamab Vedotin (RC48), Trastuzumab MMAE and Trastuzumab-DM1 conjugated antibody respectively. The coating antibody was Monoclonal Anti-MMAE&MMAF Antibody, Mouse IgG1 (Cat. No. MME-M5252), used at 1 μg/mL concentration. The primary antibody were different payload conjugated antibodies, including Trastuzumab Deruxtecan, IgG1-MMAF, Sacituzumab Govitecam, Disitamab Vedotin (RC48), Trastuzumab MMAE and Trastuzumab-DM1 conjugated antibodies used at 0.5 μg/mL concentration. The secondary antibody was Peroxidase AffiniPure Goat Anti-Human IgG, Fcγ fragment specific (min X Bov, Hrs, Ms Sr Prot) (Jackson, Cat. No. 109-035-098) used at 1:12000 concentration. Monoclonal Anti-MMAE&MMAF Antibody, Mouse IgG1 (Cat. No. MME-M5252) is specific to Disitamab Vedotin (RC48), Trastuzumab MMAE and IgG1-MMAF, and has no cross-reactivity with Trastuzumab Deruxtecan, Sacituzumab Govitecam and Trastuzumab-DM1 (Routinely tested).

Serial dilutions of Monomethyl auristatin E were added into Monoclonal Anti-MMAE specific Antibody, Rabbit IgG (M1H05) (Cat. No. MME-MY2198a.): Disitamab Vedotin (RC48) binding reactions. The half maximal inhibitory concentration (IC50) is 2.642 μg/mL (Routinely tested).