Antibody-drug Conjugate Information
General Information of This Antibody-drug Conjugate (ADC)
| ADC ID |
DRG0JFYCH
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| ADC Name |
KK2845
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| Synonyms |
KK2845
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| Organization |
Kyowa Kirin (Originator)
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| Drug Status |
Phase 1
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| Structure |
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| Antibody Name |
TIM-3Ab
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Antibody Info | ||||
| Antigen Name |
T-cell immunoglobulin mucin-3 (TIM-3)
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Antigen Info | ||||
| Payload Name |
SG3199 (SC-DR002)
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Payload Info | ||||
| Therapeutic Target |
Human deoxyribonucleic acid (hDNA)
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Target Info | ||||
| Linker Name |
Mal-PEG8-Val-Ala-PABC
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Linker Info | ||||
| Conjugate Type |
Random Cysteines
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| Combination Type |
tesirine
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The indication landscape of This ADC(2027 Update)
The clinical trial pipeline of This ADC(2027 Update)
| Indication | Phase 1 | Phase 1/2 | Phase 2 | Phase 2/3 | Phase 3 | New Drug Application | Approved | ||
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| Acute myeloid leukaemia |
1 Trials
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ADC-specific functional property(2027 Update)
Binding Affinity
| Dissocation Constant (Kd) | Binding Target | Description | Reference |
|---|---|---|---|
| 1.64 nM |
Human TIM-3
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Binding Affinity of KK2845 against human, cynomolgus monkey, mouse, rat, and rabbit TIM-3.
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[1]
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| 6.09 nM |
Cynomolgus monkey TIM-3
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Binding Affinity of KK2845 against human, cynomolgus monkey, mouse, rat, and rabbit TIM-3.
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[1]
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General Information of The ADMET Data Related to This ADC(2027 Update)
Absorption
| Standard Type | Value | Units | Description | Reference |
|---|---|---|---|---|
| Maximum Observed Concentration (Cmax) | 6.2 | ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, Male.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 12.6 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, Male, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 13 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, Male, AUCinf.
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[1] |
| Maximum Observed Concentration (Cmax) | 5.51 | ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, female.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 11.7 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, female, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 12.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, female, AUCinf.
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[1] |
| Maximum Observed Concentration (Cmax) | 12.7 | ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, Male.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 31.9 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, Male, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 33.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, Male, AUCinf.
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[1] |
| Maximum Observed Concentration (Cmax) | 12.8 | ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, female.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 24.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, female, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 26.2 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, female, AUCinf.
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[1] |
| Maximum Observed Concentration (Cmax) | 18.9 | ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, Male.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 52 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, Male, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 54.3 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, Male, AUCinf.
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[1] |
| Maximum Observed Concentration (Cmax) | 17.8 | ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, female.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 31.8 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, female, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 32.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, female, AUCinf.
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[1] |
Distribution
| Standard Type | Value | Units | Description | Reference |
|---|---|---|---|---|
| Area Under the Concentration-Time Curve (AUC) | 12.6 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, Male, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 13 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, Male, AUCinf.
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[1] |
| Volume of Distribution (Vd) | 50.2 | mL/kg |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, Male.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 11.7 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, female, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 12.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, female, AUCinf.
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[1] |
| Volume of Distribution (Vd) | 54 | mL/kg |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, female.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 31.9 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, Male, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 33.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, Male, AUCinf.
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[1] |
| Volume of Distribution (Vd) | 60.9 | mL/kg |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, Male.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 24.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, female, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 26.2 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, female, AUCinf.
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[1] |
| Volume of Distribution (Vd) | 53.2 | mL/kg |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, female.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 52 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, Male, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 54.3 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, Male, AUCinf.
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[1] |
| Volume of Distribution (Vd) | 60.3 | mL/kg |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, Male.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 31.8 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, female, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 32.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, female, AUCinf.
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[1] |
| Volume of Distribution (Vd) | 58 | mL/kg |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, female.
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[1] |
Metabolism
| Standard Type | Value | Units | Description | Reference |
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| Area Under the Concentration-Time Curve (AUC) | 12.6 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, Male, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 13 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, Male, AUCinf.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 11.7 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, female, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 12.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, female, AUCinf.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 31.9 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, Male, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 33.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, Male, AUCinf.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 24.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, female, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 26.2 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, female, AUCinf.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 52 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, Male, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 54.3 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, Male, AUCinf.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 31.8 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, female, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 32.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, female, AUCinf.
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[1] |
Excretion
| Standard Type | Value | Units | Description | Reference |
|---|---|---|---|---|
| Elimination Half-Life (t1/2) | 3.33 | day |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, Male.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 12.6 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, Male, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 13 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, Male, AUCinf.
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[1] |
| Clearance (CL) | 15.7 | mL/day/kg |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, Male.
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[1] |
| Elimination Half-Life (t1/2) | 2.68 | day |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, female.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 11.7 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, female, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 12.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, female, AUCinf.
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[1] |
| Clearance (CL) | 16.3 | mL/day/kg |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.2 mg/kg, female.
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[1] |
| Elimination Half-Life (t1/2) | 4.6 | day |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, Male.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 31.9 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, Male, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 33.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, Male, AUCinf.
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[1] |
| Clearance (CL) | 12.5 | mL/day/kg |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, Male.
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[1] |
| Elimination Half-Life (t1/2) | 3.31 | day |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, female.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 24.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, female, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 26.2 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, female, AUCinf.
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[1] |
| Clearance (CL) | 16.3 | mL/day/kg |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.4 mg/kg, female.
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[1] |
| Elimination Half-Life (t1/2) | 5.01 | day |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, Male.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 52 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, Male, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 54.3 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, Male, AUCinf.
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[1] |
| Clearance (CL) | 11.6 | mL/day/kg |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, Male.
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[1] |
| Elimination Half-Life (t1/2) | 3.27 | day |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, female.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 31.8 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, female, AUCtau.
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[1] |
| Area Under the Concentration-Time Curve (AUC) | 32.4 | day*ug/mL |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, female, AUCinf.
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[1] |
| Clearance (CL) | 19.9 | mL/day/kg |
The concentrations of the conjugated antibody (antibody with at least one payload conjugated) in cynomolgus monkeys, 0.6 mg/kg, female.
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[1] |
General Information of The Activity Data Related to This ADC
Revealed Based on the Cell Line Data
Full List of Activity Data of This Antibody-drug Conjugate
Revealed Based on the Cell Line Data
| Experiment 1 Reporting the Activity Date of This ADC | [1] | ||||
| Efficacy Data | Half Maximal inhibitory Concentration (lC50) | 1.09 ng/mL | High TIM-3 expression (TIM-3 +++) | ||
| Method Description |
EoL-1, EoL-1/hTIM-3 and Kasumi-3 cells were incubated with increasing concentrations of KK2845 or KM8047-SG3249 in RPMI1640 medium supplemented with 10 vol% (for EoL-1 and EoL-1/hTIM-3 cells) or 20 vol% (for Kasumi-3 cells) heat-inactivated fetal bovine serum, 20 vol% heat-inactivated human serum (Sigma), and a mixture of 100 U/mL penicillin and 100 ug/mL streptomycin at 37 °C for 4 days. After incubation, the cells were treated with Cell Counting Kit-8 (CCK-8, Dojindo Laboratories), and the absorbance was measured using a Multiskan Ascent device (Thermo Fisher Scientific, Ascent Software: version 2.6). Primary AML cells were incubated with increasing concentrations of KM8047-SG3249, KK2845, or CD33-ADC in IMDM medium (Thermo Fisher Scientific) supplemented with 20 vol% heat-inactivated human serum, a mixture of 100 U/mL penicillin and 100 ug/mL streptomycin, 25 ng/mL hIL-3 (Miltenyi Biotec), 25 ng/mL hSCF (Peprotech), 25 ng/mL hGM-CSF (R&D), and 25 ng/mL hG-CSF (R&D) at 37 °C for 4 days. After incubation, the cells were stained by anti-CD45 antibody-FITC (BD Pharmingen, HI30 clone), anti-CD34 antibody-APC (BD Bioscience, 8G12 clone), anti-CD38 antibody-BV421 (BD Bioscience, HIT2 clone) and 7-amino-actinomycin D (7-AAD, Bio-Rad Laboratories). The number of viable cells in the CD34+ fraction, CD34+CD38- fraction, or CD34- fraction in primary AML cells was counted by flow cytometry (FACSVerse, software program: BD FACSUITE-V1.0.5).
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| In Vitro Model | Chronic eosinophilic leukemia, not otherwise specified,Chronic eosinophilic leukemia | EoL-1/hTIM-3 cells | CVCL_0258 | ||
| Experiment 2 Reporting the Activity Date of This ADC | [1] | ||||
| Efficacy Data | Half Maximal inhibitory Concentration (lC50) | 49.58 ng/mL | Low TIM-3 expression (TIM-3+) | ||
| Method Description |
EoL-1, EoL-1/hTIM-3 and Kasumi-3 cells were incubated with increasing concentrations of KK2845 or KM8047-SG3249 in RPMI1640 medium supplemented with 10 vol% (for EoL-1 and EoL-1/hTIM-3 cells) or 20 vol% (for Kasumi-3 cells) heat-inactivated fetal bovine serum, 20 vol% heat-inactivated human serum (Sigma), and a mixture of 100 U/mL penicillin and 100 ug/mL streptomycin at 37 °C for 4 days. After incubation, the cells were treated with Cell Counting Kit-8 (CCK-8, Dojindo Laboratories), and the absorbance was measured using a Multiskan Ascent device (Thermo Fisher Scientific, Ascent Software: version 2.6). Primary AML cells were incubated with increasing concentrations of KM8047-SG3249, KK2845, or CD33-ADC in IMDM medium (Thermo Fisher Scientific) supplemented with 20 vol% heat-inactivated human serum, a mixture of 100 U/mL penicillin and 100 ug/mL streptomycin, 25 ng/mL hIL-3 (Miltenyi Biotec), 25 ng/mL hSCF (Peprotech), 25 ng/mL hGM-CSF (R&D), and 25 ng/mL hG-CSF (R&D) at 37 °C for 4 days. After incubation, the cells were stained by anti-CD45 antibody-FITC (BD Pharmingen, HI30 clone), anti-CD34 antibody-APC (BD Bioscience, 8G12 clone), anti-CD38 antibody-BV421 (BD Bioscience, HIT2 clone) and 7-amino-actinomycin D (7-AAD, Bio-Rad Laboratories). The number of viable cells in the CD34+ fraction, CD34+CD38- fraction, or CD34- fraction in primary AML cells was counted by flow cytometry (FACSVerse, software program: BD FACSUITE-V1.0.5).
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| In Vitro Model | Adult acute myeloid leukemia | Kasumi-3 cells | CVCL_0612 | ||
| Experiment 3 Reporting the Activity Date of This ADC | [1] | ||||
| Efficacy Data | Half Maximal inhibitory Concentration (lC50) | 553.7 ng/mL | Low TIM-3 expression (TIM-3+) | ||
| Method Description |
EoL-1, EoL-1/hTIM-3 and Kasumi-3 cells were incubated with increasing concentrations of KK2845 or KM8047-SG3249 in RPMI1640 medium supplemented with 10 vol% (for EoL-1 and EoL-1/hTIM-3 cells) or 20 vol% (for Kasumi-3 cells) heat-inactivated fetal bovine serum, 20 vol% heat-inactivated human serum (Sigma), and a mixture of 100 U/mL penicillin and 100 ug/mL streptomycin at 37 °C for 4 days. After incubation, the cells were treated with Cell Counting Kit-8 (CCK-8, Dojindo Laboratories), and the absorbance was measured using a Multiskan Ascent device (Thermo Fisher Scientific, Ascent Software: version 2.6). Primary AML cells were incubated with increasing concentrations of KM8047-SG3249, KK2845, or CD33-ADC in IMDM medium (Thermo Fisher Scientific) supplemented with 20 vol% heat-inactivated human serum, a mixture of 100 U/mL penicillin and 100 ug/mL streptomycin, 25 ng/mL hIL-3 (Miltenyi Biotec), 25 ng/mL hSCF (Peprotech), 25 ng/mL hGM-CSF (R&D), and 25 ng/mL hG-CSF (R&D) at 37 °C for 4 days. After incubation, the cells were stained by anti-CD45 antibody-FITC (BD Pharmingen, HI30 clone), anti-CD34 antibody-APC (BD Bioscience, 8G12 clone), anti-CD38 antibody-BV421 (BD Bioscience, HIT2 clone) and 7-amino-actinomycin D (7-AAD, Bio-Rad Laboratories). The number of viable cells in the CD34+ fraction, CD34+CD38- fraction, or CD34- fraction in primary AML cells was counted by flow cytometry (FACSVerse, software program: BD FACSUITE-V1.0.5).
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| In Vitro Model | Chronic eosinophilic leukemia | EoL-1 cells | CVCL_0258 | ||
References