1. Supplementary Figure 1: AGS-22M6E ADC specific localization in tumor xenografts. AG-B1 tumor pieces were implanted in ICR-SCID mice subcutaneously. When tumors reached approximately 200 mm3 in volume, mice were untreated (A), treated intravenously with 10 mg/kg isotype control ADC (B, C, D) or treated with AGS-22M6E (E, F, G). Mice were sacrificed at the indicated time points. FFPE specimens were prepared from excised tumors. FFPE sections were stained with the anti-MMAE antibody SG15-22 and detection was performed using the Biogenex Super Sensitive Polymer-HRP IHC Detection Kit.

2. Non-treated A.
Control ADC treated
AGS-22M6E treated
6hrs B. E.
24 hrs C. F.
72 hrs D. G.

3. Supplementary Figure 2: Serum Concentration time profiles for AGS-22M6E and ASG-22CE (A) ADC and (B) Total IgG. AGS-22M6E and ASG-22CE were administered via single intravenous bolus injection into ICR SCID mice at a dose of 10 mg/kg. Sera was collected from whole blood at various time points (n=4/group). A quantitative sandwich enzyme linked immunoassay (ELISA) was used to measure the amount of total IgG antibody and the amount of ADC in the serum samples by capturing with an anti-idiotype antibody and anti-MMAE antibody, respectively.

4. A. ADC
B. Total IgG

5. Supplementary Table 1: Non Compartmental Toxicokinetic parameters in ICR SCID mice following a single IV bolus injection of AGS-22M6E and ASG-22CE at 10 mg/kg.
Assay
Antibody
T1/2
(day)
Cmax
(µg/mL)
AUClast (day*µg/mL)
AUCinf
(day*µg/mL)
Vss
(mL/kg) CL
(mL/day/kg)
ADC
AGS-22M6E
1.53
135
336
339
78.1
29.4
ASG-22CE
1.72
177
324
329
82.4
30.3
Total IgG
1.73
148
363
370
82.1
27.0
2.08
200
349
87.1
28.6
Mean ADC and Total IgG serum concentration obtained following single dose was analyzed by non-compartmental methods using WinNonlin Pro version (Pharsight Corp., Mountain View, CA).
T1/2: elimination half-life; Cmax: maximum concentration observed; AUClast: Area under the curve from time zero to last observable time point; AUCinf: Area under the curve from time zero to infinity; Vss: volume of distribution at steady state; CL: clearance