Antibody-drug Conjugate Information

General Information of This Antibody-drug Conjugate (ADC)
ADC ID
DRG0ZXHDJ
ADC Name
aTCRA6 DAR2-8
Synonyms
aTCRA6 DAR2-8
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Organization
Technical University of Darmstadt; Goethe University; Frankfurt Cancer Institute; German Cancer Consortium (DKTK); German Cancer Research Center (DKFZ).
Drug Status
Investigative
Drug-to-Antibody Ratio
5
Structure
Antibody Name
aTCRA6S1
  Antibody Info 
Antigen Name
Transcriptional regulatory protein TcrA (TCRA6)
  Antigen Info 
Payload Name
Monomethyl auristatin E
  Payload Info 
Therapeutic Target
Microtubule (MT)
  Target Info 
Linker Name
DBCO-PEG4-Val-Cit-PABC
  Linker Info 
Conjugate Type
Random conjugation through reduced inter-chain cysteines.
ADC-specific functional property(2027 Update)
Bystander Killing Effect
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Bystander Killing Effect Description Reference
yes
Circumventing such tumor escape mechanisms, an ADC is capable to mediate effects, (mainly) by internalization and degradation in target cells and subsequent permeation and diffusion of free payload, leading to the death of surrounding cells. Investigating the in vitro activity of our aTCRA6 DAR2 and aTCRA6 DAR52-8 ADCs, direct cell killing was measured using Jurkat TCRA6 target cells, and killing was analyzed by transfer of respective cell culture supernatants to Jurkat WT off-target cells. Consistent with our preliminary cytotoxicity data, direct killing of TCRA6-positive Jurkat cells was potently induced by the ADCs post 96 h.

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[1]
General Information of The Activity Data Related to This ADC
Revealed Based on the Cell Line Data
Click To Hide/Show 2 Activity Data Related to This Level
Standard Type Value Units Cell Line Disease Model
Half Maximal Effective Concentration (EC50) 
0.1
nM
CVCL_0065
Childhood T acute lymphoblastic leukemia, Precursor T-cell acute lymphoblastic leukemia
Half Maximal Effective Concentration (EC50) 
> 100
nM
CVCL_0065
Childhood T acute lymphoblastic leukemia, Precursor T-cell acute lymphoblastic leukemia
Full List of Activity Data of This Antibody-drug Conjugate
Revealed Based on the Cell Line Data
Click To Hide/Show 2 Activity Data Related to This Level
Experiment 1 Reporting the Activity Date of This ADC [1]
Efficacy Data Half Maximal Effective Concentration (EC50) 0.1 nM Positive TCRA6 expression (TCRA6+++/++)
Method Description
Cytotoxic effects of aTCRA6 DAR2 and aTCRA6 DAR52-8 ADCs were estimated by exposing Jurkat WT and Jurkat TCRA6 T cells to different ADC concentrations. Cell viability was analyzed 72 h post ADC addition by a fluorometric method using CellTiter-Blue Cell Viability Assay (Promega, Wisconsin, USA) according to the manufacturer's instructions. Briefly, cells were seeded in a 96-well plate (1 × 104 cells/well) with the desired ADC concentrations ranging from 0.06-400 nM (for DAR2 ADC) or 0.005-30 nM (for DAR52-8 ADC) in a serial dilution. After 72 h, redox dye (resazurin) was added to the cells, and the plate was incubated for 3-4 h. Fluorescence of resorufin (560Ex/590Em) was recorded using CLARIOstar plus microplate reader (BMG LABTECH, Offenburg, Germany). Cell proliferation was normalized to untreated control cell fluorescence values. The resulting curves were fitted with a variable slope four-parameter fit, and EC50s were calculated using GraphPad Prism.

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In Vitro Model Childhood T acute lymphoblastic leukemia, Precursor T-cell acute lymphoblastic leukemia Jurkat cells (TCRA6+) CVCL_0065
Experiment 2 Reporting the Activity Date of This ADC [1]
Efficacy Data Half Maximal Effective Concentration (EC50) > 100 nM Negative TCRA6 expression (TCRA6-)
Method Description
Cytotoxic effects of aTCRA6 DAR2 and aTCRA6 DAR52-8 ADCs were estimated by exposing Jurkat WT and Jurkat TCRA6 T cells to different ADC concentrations. Cell viability was analyzed 72 h post ADC addition by a fluorometric method using CellTiter-Blue Cell Viability Assay (Promega, Wisconsin, USA) according to the manufacturer's instructions. Briefly, cells were seeded in a 96-well plate (1 × 104 cells/well) with the desired ADC concentrations ranging from 0.06-400 nM (for DAR2 ADC) or 0.005-30 nM (for DAR52-8 ADC) in a serial dilution. After 72 h, redox dye (resazurin) was added to the cells, and the plate was incubated for 3-4 h. Fluorescence of resorufin (560Ex/590Em) was recorded using CLARIOstar plus microplate reader (BMG LABTECH, Offenburg, Germany). Cell proliferation was normalized to untreated control cell fluorescence values. The resulting curves were fitted with a variable slope four-parameter fit, and EC50s were calculated using GraphPad Prism.

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In Vitro Model Childhood T acute lymphoblastic leukemia, Precursor T-cell acute lymphoblastic leukemia Jurkat cells (TCRA6-) CVCL_0065
References
Ref 1 T cell receptor-directed antibody-drug conjugates for the treatment of T cell-derived cancers