1. Supplementary Table 1. TTI-621 binds to human tumor cell lines
Tumor Type
Cell Line
Binding EC50 (nM)
Acute myeloid leukemia
OCI-AML-2
262
B cell lymphoma
Pfeiffer
493 HT 20
SU8 16
Toledo 23
SU4 90
SU6 32
NU1 31
SU16 14
Breast cancer
SKBR3
461
Epidermoid carcinoma
A431
136
Multiple myeloma
H929
253
MM1s
171
T cell leukemia
Jurkat
T cell lymphoma HH 66 H9 34
Supplementary Table 1: Established human tumor cell lines were incubated with titrated amounts of biotinylated TTI-621, washed and stained with streptavidin-PE. Cells were then washed, fixed and analyzed by flow cytometry. EC50 values were generated by linear regression using a sigmoidal dose- response curve.

2. Supplementary Table 2. TTI-621 binds to human primary tumor samples
Tumor Type
Patient ID
Binding EC50 (nM)
Acute myeloid leukemia A1
161 A2
333
B-acute lymphoblastic leukemia B1 72 B2 80 B3
154 B4
106 B5
164 B6
109 B7 75 B8
299 B9
425
B10
807
Myelodysplastic syndrome M1 93 M2 83
T-acute lymphoblastic leukemia T1 77 T2 81 T3 97 T4
346 T5
176
Supplementary Table 2: Primary human tumor samples were incubated with titrated amounts of biotinylated TTI-621, washed and stained with streptavidin-PE. Cells were then washed, fixed and analyzed by flow cytometry. EC50 values were generated by linear regression using a sigmoidal dose-response curve.

3. Supplementary Table 3. TTI-621 binds to peripheral blood cells of healthy donors
Cell type
Binding EC50 (nM)
CD4+ T cells
153
CD8+ T cells
168
B cells
123
NK cells
902
NKT cells
293
Platelets 57
Supplementary Table 3: Fresh heparinized whole blood from healthy donors (n=6) was erythrocyte-lysed. Cells were incubated with titrated amounts of biotinylated TTI-621 (0.24 nM-10 μM) or control IgG1 Fc, washed, and stained with streptavidin-PE. Cells were then washed, fixed and analyzed by flow cytometry. Cells were gated as live, single cells and defined as follows: B cells: CD3-CD20+, CD4+ T cells: CD3+CD4+, CD8+ T cells: CD3+CD4-, NK cells: CD3-CD56+, NKT cells: CD3+CD56+, platelets: CD45-CD41b+. EC50 values were generated by linear regression using a sigmoidal dose-response curve.

4. Supplementary Table 4: TTI-621 Potency In Vitro
Tumor Type
Sample
Phagocytosis EC50 (nM)
Acute myeloid leukemia
OCI-AML-2 cell line 5
Patient sample A3 4
Patient sample A4 2
Patient sample A5 57
B cell lymphoma
Pfeiffer cell line 8
Raji cell line 3
SUDHL6 cell line
0.5
SUDHL16 cell line 16
Toledo cell line
Breast cancer
SKBR3 cell line 23
Lung cancer
A549 cell line
Melanoma
A375.S2 cell line
Multiple myeloma
MM.1s cell line 6
H929 cell line
Patient sample M3
T cell leukemia
Jurkat cell line
0.08
Supplementary Table 4: Phagocytosis of established human tumor cell lines and primary human tumor samples was assessed in the presence of titrated amounts of TTI-621. EC50 values were generated by linear regression using a sigmoidal dose-response curve.