Anti-cd33 and Anti-cd7 combination treatment
An antigen and antibody technology, applied in the direction of drug combination, antibody, anti-receptor/cell surface antigen/cell surface determinant immunoglobulin, etc., can solve life-threatening infection, bone marrow cell consumption and other problems
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Embodiment 1
[0194] Example 1: Determination of IC of 5 monoclonal antibodies (individually and in combination) in Kasumi-3 cell line 50
[0195] Experimental conditions
[0196] Table A: Overview of experimental conditions tested.
[0197]
[0198] Note: CD(A)=CD33; CD(B)=CD7; CD(C)=CD13
[0199] Number of test samples
[0200] 5 antibodies or combinations * 8 concentrations * 1 cell line * 3 (triplicates) = 120
[0201] Negative control (anti-Fc linked to MMAE) * 1 concentration * 1 cell line * 3 (triplicate) = 3
[0202] 3 positive controls (primary antibodies) * 1 concentration * 1 cell line * 3 (triplicates) = 9
[0203] Control (cells without antibody treatment) * 1 cell line * 3 (triplicates) = 3
[0204] Total = 135 tests
[0205] 2.Material:
[0206] i.Kasumi-3: CRL-2725 TM
[0207] ii. RPMI-1640 medium: 30-2001 TM
[0208] iii. Fetal Bovine Serum (FBS): 30-2020 TM
[0209] iv. Dulbecco's Phosphate Buffered Saline (DPBS): Gibco TM
[0210] v. Monoclona...
Embodiment 2
[0276] Example 2: Determination of IC of 3 monoclonal antibodies (alone and in combination) in the dual antigen cell line HEL 92.1.7 50
[0277] Experimental conditions
[0278] Table C: Overview of experimental conditions tested
[0279]
[0280] Note: CD(A) = CD33; 1; CD(D) = CD56
[0281] Number of test samples
[0282] - 3 antibodies or combinations * 8 concentrations * 1 cell line * 3 (triplicates) = 72
[0283] - Negative control (anti-Fc linked to MMAE) * 1 concentration * 1 cell line * 3 (triplicates) = 3
[0284] - 2 positive controls (primary antibodies) * 1 concentration * 1 cell line * 3 (triplicates) = 6
[0285] - Control (cells not treated with antibody) * 1 cell line * 3 (triplicates) = 3
[0286] Total = 84 tests
[0287] 2.Material:
[0288] i.HEL 92.1.7: TIB-180
[0289] ii. RPMI-1640 medium: 30-2001 TM
[0290] iii. Fetal Bovine Serum (FBS): 30-2020 TM
[0291] iv. Dulbecco's Phosphate Buffered Saline (DPBS): Gibco TM
[0292] v. M...
Embodiment 3
[0347] Example 3: The second cell killing assay was repeated for Kasumi-3 using BiFab
[0348] Experiments were performed against the Kasumi-3 cell line using BiFab to repeat the second cell killing assay.
[0349] reagent
[0350]
[0351] growth medium
[0352] RPMI-1640 Medium
[0353] 10% Fetal Bovine Serum
[0354] 1% GlutaMAX Supplement
[0355] method
[0356] Harvest Kasumi-3 cells, count and divide at 2x10 4 Cells / well were seeded into 100 μl of growth medium in a 96-well plate. A 5-point dose response of WT Bifab was prepared in growth medium at 50-fold assay final concentration. 2 μl of WT Bifab titer was pipetted onto the seeded cells and each concentration was determined on triplicate wells. Cells were incubated at room temperature for 10 minutes to allow Bifab to bind. At the same time, the anti-human IgG Fab-MMAE antibody was diluted in growth medium to provide an assay final concentration of 6 nM MMAE / well, as described in the manufacturer's...
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