KEY RESOURCES TABLE
| REAGENT or RESOURCE | SOURCE | IDENTIFIER |
|---|---|---|
| Antibodies | ||
| N88A/N297A anti-EGFR mAb | This paper | N/A |
| N297A anti-EGFRvIII mAb | This paper | N/A |
| N297Q anti-EGFRvIII mAb | This paper | N/A |
| N297A anti-HER2 mAb | This paper | N/A |
| Rabbit anti-MMAF antibody | Levena Biopharma | Cat#LEV-PAF1 |
| Goat anti-human IgG Fab-horseradish peroxidase (HRP) conjugate | Jackson ImmunoResearch | Cat#109-035-097; RRID:AB_2337585 |
| Goat anti-human IgG Fc antibody | Jackson ImmunoResearch | Cat#109-005-098; RRID:AB_2337541 |
| Donkey anti-human IgG-HRP conjugate | Jackson ImmunoResearch | Cat#709-035-149; RRID:AB_2340495 |
| Goat anti-rabbit IgG–HRP conjugate | Thermo Fisher Scientific | Cat#32260; RRID:AB_1965959 |
| Rabbit anti-cleaved caspase 3 antibody | Cell Signaling Technology | Cat#9661; RRID:AB_2341188 |
| Rabbit anti-EGFR antibody | Cell Signaling Technology | Cat#4267; RRID:AB_2246311 |
| Rabbit anti-Ki67 antibody | Abcam | Cat#Ab16667; RRID:AB_302459 |
| Bacterial and virus strains | ||
| HLUC-Lv105 Firefly Luciferase Lentifect™ Purified Lentiviral Particles | GeneCopoeia | Cat#LP461-025 |
| pLL-CMV-RFP-T2A-Puro (Lenti-Labeler™ virus) | System Biosciences | Cat#LL110VA-1 |
| Chemicals, peptides, and recombinant proteins | ||
| Monomethyl auristatin F (MMAF) | Levena Biopharma | Cat# T1006 |
| diazide branched linker | Anami et al., 2018 | https://doi.org/10.1038/s41467-018-04982-3 |
| BCN–EVCit–PABC–MMAF | This paper | N/A |
| MC–MMAF | This paper | CAS 863971-19-1 |
| MMAF-NHS | This paper | N/A |
| DBCO–EVCit–Cy5.5 | This paper | N/A |
| BCN-peg4-MMAF | This paper | N/A |
| TCO-peg4-MMAF | This paper | N/A |
| Click-easy™ BCN N-hydroxysuccinimide Ester | Berry & Associates | Cat#LK4320 |
| TCO-NHS ester | BroadPharm | Cat#BP-22417 |
| Sulfo-Cyanine5.5 amine (Cy5.5-amine) | Lumiprobe | Cat#273C0 |
| Cy5.5 maleimide | Click Chemistry Tools | Cat#1113-1 |
| Cy5.5-NHS ester | Click Chemistry Tools | Cat#1056-1 |
| Activa TI® | Modernist Pantry | Cat#1002 |
| Human EGFR Protein, His Tag (MALS verified) | ACROBiosystems | Cat#EGR-H5222 |
| DAPI | VECTOR | Cat#H-1200 |
| FITC-conjugated dextran | Sigma-Aldrich | Cat#FD500S |
| Homogeneous anti-EGFR ADC 1 | This paper | N/A |
| Cys conjugate 2 | This paper | N/A |
| Lys conjugate 3 | This paper | N/A |
| Homogeneous anti-EGFRvIII ADC 4 | This paper | N/A |
| Heterogeneous anti-EGFRvIII ADC 5 | This paper | N/A |
| Homogeneous anti-HER2 ADC 6 | This paper | N/A |
| Heterogeneous anti-HER2 ADC 7 | This paper | N/A |
| Homogeneous Cy5.5 conjugate 8 | This paper | N/A |
| Cys-Cy5.5 conjugate 9 | This paper | N/A |
| Lys-Cy5.5 conjugate 10 | This paper | N/A |
| Homogeneous Cy5.5 conjugate (non-cleavable) | This paper | N/A |
| DAR 0 mAb-Cy5.5 conjugate 11 | This paper | N/A |
| DAR 4 ADC-Cy5.5 conjugate 12 | This paper | N/A |
| DAR 6 ADC-Cy5.5 conjugate 13 | This paper | N/A |
| DAR 8 ADC-Cy5.5 conjugate 14 | This paper | N/A |
| Critical commercial assays | ||
| BCA protein assay | Thermo Fisher Scientific | Cat#23225 |
| WST-8 | Cayman Chemical | Cat#18721 |
| 1-methoxy-5-methylphenazinium methylsulfate | Cayman Chemical | Cat#21258 |
| SignalStain® Boost IHC Detection Reagent | Cell Signaling Technology | Cat#8114P |
| SignalStain® DAB Substrate Kit | Cell Signaling Technology | Cat#8059P |
| Experimental models: Cell lines | ||
| U87ΔEGFR | Dr. Erwin G. Van Meir (Emory University) | N/A |
| U87ΔEGFR-luc | This paper | N/A |
| Gli36δEGFR | Dr. E. Antonio Chiocca (Brigham and Women’s Hospital, Harvard Medical School) | N/A |
| HEK293 | ATCC | Cat#CRL-1573 |
| JIMT1-BR3 | Dr. Patricia S. Steeg (National Cancer Institute) | N/A |
| GBM12 | Dr. Jann N. Sarkaria (Mayo Clinic) | N/A |
| RFP-expressing GBM12 (GBM12-RFP) | This paper | N/A |
| FreeStyle™ 293-F Cells | Thermo Fisher Scientific | Cat#R79007 |
| Experimental models: Organisms/strains | ||
| NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ mouse | The Jackson Laboratory | Cat#005557 |
| CD-1® IGS Mouse | Charles River Laboratories | Cat#022 |
| Software and algorithms | ||
| GraphPad Prism 8 | GraphPad Software | https://www.graphpad.com/scientific-software/prism/ |
| Gen5 | Agilent | https://www.biotek.com/products/software-robotics-software/gen5-microplate-reader-and-imager-software/ |
| NIS Elements AR software | Nikon | https://www.microscope.healthcare.nikon.com/products/software/nis-elements |
| Microsoft Excel 16 | Microsoft | https://www.microsoft.com/en-us/download/details.aspx?id=56547 |
| Image J | NIH | https://imagej.nih.gov/ij/download.html |
| PKSolver version 2 | Zhang et al., 2010 | https://doi.org/10.1016/j.cmpb.2010.01.007 |
| LI-COR Image Studio | LI-COR | https://www.licor.com/bio/image-studio/ |