Streptavidin-ZAP References

Cat #IT-27


Yuan X, Yang M, Chen X, Zhang X, Sukhadia S, Musolino N, Bao H, Chen T, Xu C, Wang Q, Santoro S, Ricklin D, Hu J, Lin R, Yang W, Li Z, Qin W, Zhao A, Scholler N, & Coukos G. Characterization of the first fully human anti-TEM1 scFv in models of solid tumor imaging and immunotoxin-based therapy. (2018). Cancer Immunol Immunother, 67 (2):329-339. 2018/01/10.

Objective: To evaluate potential applications of scFv78 as a tool for tumor molecular imaging, immunotoxin-based therapy and nanotherapy.
Summary: The data indicate that scFv78 has unique and favorable features that are advantageous for the development of imaging probes or antibody-toxin conjugates for a large spectrum of human TEM1-positive solid tumors.
Dose: Biotinylated ScFv78 was conjugated with Streptavidin-ZAP at a molar ratio of 4:1 (scFv78:ZAP).

Lelieveldt LPWM, Kristyanto H, Pruijn GJM, Scherer HU, Toes REM, & Bonger KM. Sequential Prodrug Strategy to Target and Eliminate ACPA-Selective Autoreactive B cells. (2018). Mol Pharm, Epub 26 Oct 2018

Objective: To develop a method to target and selectively eliminate autoreactive B cells using a sequential antigen prodrug targeting strategy.
Summary:  The selectivity of the antigen and the possibility to block binding toward circulation ACPA brings us a step closer to the specific elimination of autoreactive B cells for the treatment of patients with ACPA-positive RA.
Dose:  Biotinylated CCP1, CArgP1, and CCP1(CNBz) were conjugated with Streptavidin-ZAP in a 4:1 ratio to make peptide-drug conjugates.

Berhani O, Glasner A, Kahlon S, Duev-Cohen A, Yamin R, Horwitz E, Enk J, Moshel O, Varvak A, Porgador A, Jonjic S, & Mandelboim O. Human anti-NKp46 antibody for studies of NKp46-dependent NK cell function and its applications for type 1 diabetes and cancer research. (2018). Eur J Immunol (epub ahead of print)

Objective:  To investigate human NKp46 activity and its critical role in Natural Killer (NK) cell biology.
Summary:  A unique anti-human NKp46 monoclocal antibody was developed and conjugated to Saporin.  Targeted toxin inhibits growth of NKp46-positive cells; thus, exemplifying the potential as an immunotherapeutic drug to treat NKp46-dependent diseases, such as, type I diabetes and NK and T cell related malignancies.
Dose:  Conjugation of the antibodies to Saporin, treatment of cells, and cell viability assay Biotin-Z Kit instructions.

Palchaudhuri R, Hyzy SL, Proctor JL, Adams HL, Pearse BR, Sarma G, Aslanian S, Gillard G, Lamothe TL, Burenkova O, Brooks ML, Gabros AD, McDonagh CF, Boitano AE, & Cooke MP. (2018). Antibody Drug Conjugates Targeted to CD45 or CD117 Enable Allogeneic Hematopoietic Stem Cell Transplantation in Animal Models. Paper presented at the ASH Annual Meeting, San Diego, CA.

Objective:  To further investigate and develop the utility of CD45-SAP and CD117-SAP, in combination with immunosuppression, in murine transplant models using i.v. administration in an allogeneic minor mismatch transplant model (Balb/c donor into DBA/2 recipients).
Summary:  CD45-SAP or CD117-SAP in combination with immunosuppressants (30Fll and post-transplant Cytoxan) enabled >85% peripheral donor chimerism at 12 weeks post-transplantation.  CD45-5AP and CD117-SAP were more effective at conditioning versus 2Gy TBI or pretransplant Cytoxan.
Dose:  CD45-SAP (1.9 mg/kg, iv) and CD117-SAP (1mg/kg, iv) in an allogeneic minor mismatch transplant model (Balb/c donor into DBA/2 recipients).

Tan HL, Yong C, Tan BZ, Fong WJ, Padmanabhan J, Chin A, Ding V, Lau A, Zheng L, Bi X, Yang Y, & Choo A. Conservation of Oncofetal Antigens on Human Embryonic Stem Cells Enables Discovery of Monoclonal Antibodies against Cancer. (2018). Scientific Reports, 8 (1):11608.

Objective:  To identify and characterize an antibody raised using human embryonic stem cells with potential as a cancer therapeutic.
Summary:  Antibody A19 not only binds to undifferentiated hESCs by flow cytometry, it also reacts with ovarian and breast cancer cell lines with low or no binding to normal cells.
DoseIn Vitro – Number of viable cells treated showed a decrease in cell number (Hum-ZAP mixed with A19; Streptavidin-ZAP mixed with biotinylated A19).  To determine if there were off-target effects, Hum-ZAP and chA19 were incubated with a non-binding cell line OVCAR10; no apparent cytotoxicity was observed.
In Vivo – 5 x 106 SKOV3 cells were implanted s.c. in NUDE mice and Biotinylated A19-Streptavidin-ZAP (ADC), administered ip.  The controls were free Saporin and naked A19.  By the end of 10 weeks, mice administered with the ADC saw a 60% reduction in tumor size compared to control groups.

Kusano-Arai O, Iwanari H, Kudo S, Kikuchi C, Yui A, Akiba H, Matsusaka K, Kaneda A, Fukayama M, Tsumoto K, & Hamakubo T. Synergistic Cytotoxic Effect on Gastric Cancer Cells of an Immunotoxin Cocktail in Which Antibodies Recognize Different Epitopes on Cdh17. (2018). Monoclonal Antibodies in Immunodiagnosis and Immunotherapy, 37 (1):1-11.

Dose:  For immunotoxin preparation, Streptavidin-ZAP was mixed with biotinylated mAbs in equal molar concentrations for 30 min at room temp.
Objective:  To determine if an immunotoxin cocktail targeted to multiple epitopes has synergistic effects on low expression level cells, which would expand the applicable range of immunotoxin therapy for cancer.
Summary:  The combination of immunotoxins with different mechanisms of action in an antibody cocktail will increase cytotoxic activities and decrease side effects.  For immunotoxin preparation, Streptavidin-ZAP was mixed with biotinylated mAbs in equal molar concentrations for 30 minutes at room temperature.


Ha KD, Bidlingmaier SM, Su Y, Lee NK, & Liu B. Identification of Novel Macropinocytosing Human Antibodies by Phage Display and High-Content Analysis. (2017). Methods Enzymol, 585 91-110. 2017/01/23. PMID: 28109445

 Streptavidin-ZAP is mixed with biotinylated IgG at a molar ratio of 1:1, vortexed, and incubated on ice for 30 min to form the immunotoxin.
Serially-diluted immunotoxin (50 mcL in PBS) is added to each well and incubated for 96 h at 37°C with 5% CO2. For initial assessment, 1:10 serial dilutions are often used to find the range. For assessment of the half-maximal effective concentration (EC50), 1:3 serial dilution is used to improve accuracy.

Schiroli G, Ferrari S, Conway A, Jacob A, Capo V, Albano L, Plati T, Castiello MC, Sanvito F, Gennery AR, Bovolenta C, Palchaudhuri R, Scadden DT, Holmes MC, Villa A, Sitia G, Lombardo A, Genovese P, Naldini L. (2017) Preclinical Modeling Highlights the Therapeutic Potential of Hematopoietic Stem Cell Gene Editing for Correction of Scid-X1. Sci Transl Med 9(411)PMID: 29021165 (read summary)

Yuan X, Yang M, Chen X, Zhang X, Sukhadia S, Musolino N, Bao H, Chen T, Xu C, Wang Q, Santoro S, Ricklin D, Hu J, Lin R, Yang W, Li Z, Qin W, Zhao A. (2017) Characterization of the First Fully Human Anti-Tem1 Scfv in Models of Solid Tumor Imaging and Immunotoxin-Based Therapy. Cancer Immunol Immunother 66(3):367-378. PMID: 27933426 (Targeting Trends 17q1)


Dickey DD, Thomas GS, Dassie JP, Giangrande PH. (2016) Method for Confirming Cytoplasmic Delivery of RNA Aptamers. Methods Mol Biol 1364:209-217. PMID: 26472453 (Targeting Trends 16q1)

Palchaudhuri R, Saez B, Hoggatt J, Schajnovitz A, Sykes DB, Tate TA, Czechowicz A, Kfoury Y, Ruchika F, Rossi DJ, Verdine GL, Mansour MK, Scadden DT. (2016) Non-genotoxic conditioning for hematopoietic stem cell transplantation using a hematopoietic-cell-specific internalizing immunotoxin. Nat Biotechnol 34(7):738-745. PMID: 27272386 (Targeting Trends 16q3)

Palchaudhuri R. (2016) Targeted depletion of hematopoietic stem cells promises safer transplantation. Targeting Trends 17(3).


Damelin M, Bankovich A, Park A, Aguilar J, Anderson W, Santaguida M, Aujay M, Fong S, Khandke K, Pulito V, Ernstoff E, Escarpe P, Bernstein J, Pysz M, Zhong W, Upeslacis E, Lucas J, Lucas J, Nichols T, Loving K, Foord O, Hampl J, Stull R, Barletta F, Falahatpisheh H, Sapra P, Gerber HP, Dylla SJ. (2015) Anti-EFNA4 Calicheamicin Conjugates Effectively Target Triple-Negative Breast and Ovarian Tumor-Initiating Cells to Result in Sustained Tumor Regressions. Clin Cancer Res 21(18):4165-4173. PMID: 26015513 (Targeting Trends 15q4)

Sims S, Klenerman P. (2015) Increasing inflationary T-cell responses following transient depletion of MCMV-specific memory T cells. Eur J Immunol 45(1):113-118. (Targeting Trends 15q1)


Alonso MN, Gregorio JG, Davidson MG, Gonzalez JC, Engleman EG. (2014) Depletion of inflammatory dendritic cells with anti-CD209 conjugated to saporin toxin. Immunol Res 58(2-3):374-377. (Targeting Trends 14q3)

Bostad M, Kausberg M, Weyergang A, Olsen CE, Berg K, Hogset A, Selbo PK. (2014) Light-Triggered, Efficient Cytosolic Release of IM7-Saporin Targeting the Putative Cancer Stem Cell Marker CD44 by Photochemical Internalization. Mol Pharm 11(8):2764-2776. (Targeting Trends 14q4)

Ehrlich D, Wang B, Lu W, Dowling P, Yuan R. (2014) Intratumoral anti-HuD immunotoxin therapy for small cell lung cancer and neuroblastoma. J Hematol Oncol 7(1):91. (Targeting Trends 15q1)

Ha KD, Bidlingmaier SM, Zhang Y, Su Y, Liu B. (2014) High-content analysis of antibody phage-display library selection outputs identifies tumor selective macropinocytosis-dependent rapidly internalizing antibodies. Mol Cell Proteomics 13(12):3320-3331. (Targeting Trends 15q2)

Lund K, Bostad M, Skarpen E, Braunagel M, Krauss S, Duncan A, Hogset A, Selbo P. (2014) The novel EpCAM-targeting monoclonal antibody 3-17I linked to saporin is highly cytotoxic after photochemical internalization in breast, pancreas and colon cancer cell lines. MAbs 6(4):1038-50.(Targeting Trends 14q2)


Burgos-Ojeda D, McLean K, Bai S, Pulaski H, Gong Y, Silva I, Skorecki K, Tzukerman M, Buckanovich RJ. (2013) A Novel Model for Evaluating Therapies Targeting Human Tumor Vasculature and Human Cancer Stem-like Cells. Cancer Res 73(12):3555-3565. (Targeting Trends 13q3)

Bostad M, Berg K, Hogset A, Skarpen E, Stenmark H, Selbo PK. (2013) Photochemical internalization (PCI) of immunotoxins targeting CD133 is specific and highly potent at femtomolar levels in cells with cancer stem cell properties. J Control Release 168(3):317-326. (Targeting Trends 13q3)

Hernandez, LI, Flenker, KS, Hernandez, FJ, Klingelhutz, AJ, McNamara, JO, 2nd, & Giangrande, PH. (2013). Methods for Evaluating Cell-Specific, Cell-Internalizing Rna Aptamers. Pharmaceuticals (Basel), 6 (3):295-319. PMID: 23894227 (read summary)

Hess SM, Young EF, Miller KR, Vincent BG, Buntzman AS, Collins EJ, Frelinger JA, Hess PR. (2013) Deletion of naive T cells recognizing the minor histocompatibility antigen HY with toxin-coupled peptide-MHC class I tetramers inhibits cognate CTL responses and alters immunodominance. Transpl Immunol 29(1-4):138-145. (Targeting Trends 14q1)

Ren C, Luan L, Wui-Man Lau B, Huang X, Yang J, Zhou Y, Wu X, Gao J, Pickard GE, So KF, Pu M. (2013) Direct Retino-Raphe Projection Alters Serotonergic Tone and Affective Behavior. Neuropsychopharmacology 38(7):1163-1175. (Targeting Trends 13q2)

Stratford EW, Bostad M, Castro R, Skarpen E, Berg K, Hogset A, Myklebost O, Selbo PK. (2013) Photochemical internalization of CD133-targeting immunotoxins efficiently depletes sarcoma cells with stem-like properties and reduces tumorigenicity. Biochim Biophys Acta 1830(8):4235-4243. (Targeting Trends 13q3)


Berstad MB, Weyergang A, Berg K. (2012) Photochemical internalization (PCI) of HER2-targeted toxins: Synergy is dependent on the treatment sequence. Biochim Biophys Acta 1820(12):1849-1858. (Targeting Trends 13q1)


Chung JS, Shiue LH, Duvic M, Pandya A, Cruz PDJ, Ariizumi K. (2011) Sezary syndrome cells overexpress syndecan-4 bearing distinct heparan sulfate moieties that suppress T-cell activation by binding DC-HIL and trapping TGF-{beta} on the cell surface. Blood 117(12):3382-3390. (Targeting Trends 11q2)

Yang MY, Chaudhary A, Seaman S, Dunty J, Stevens J, Elzarrad MK, Frankel AE, St Croix B (2011) The Cell Surface Structure of Tumor Endothelial Marker 8 (TEM8) is Regulated by the Actin Cytoskeleton. Biochim Biophys Acta 1813(1):39-49. (Targeting Trends 11q1)


Akiyoshi H, Chung JS, Tomihari M, Cruz PD, Jr., Ariizumi K (2010) Depleting Syndecan-4+ T Lymphocytes Using Toxin-Bearing Dendritic Cell-Associated Heparan Sulfate Proteoglycan-Dependent Integrin Ligand: A New Opportunity for Treating Activated T Cell-Driven Disease. J Immunol 184:3554-3561. (Targeting Trends 10q2)

Ariizumi K, Akiyoshi H, Chung J-S, Tomiharu M, Cruz Jr PD. (2010) Depletion of syndecan-4+ T lymphocytes by saporin-conjugated DC-HIL alleviates T cell-mediated imflammatory disease. Targeting Trends 11(2).

Berg K, Weyergang A, Prasmickaite L, Bonsted A, Hogset A, Strand MT, Wagner E, Selbo PK (2010) Photochemical internalization (PCI): a technology for drug delivery. Methods Mol Biol 635:133-145. (Targeting Trends 10q4)

Kuroda K, Liu H, Kim S, Guo M, Navarro V, Bander NH (2010) Saporin toxin-conjugated monoclonal antibody targeting prostate-specific membrane antigen has potent anticancer activity. Prostate 70(12):1286-1294. (Targeting Trends 10q4)

Vincent BG, Young EF, Buntzman AS, Stevens R, Kepler TB, Tisch RM, Frelinger JA, Hess PR (2010) Toxin-Coupled MHC Class I Tetramers Can Specifically Ablate Autoreactive CD8+ T Cells and Delay Diabetes in Nonobese Diabetic Mice. J Immunol 184(8):4196-4204. (Targeting Trends 10q2)


Hess PR, Buntzman AS, Murray SL, Young EF, Frelinger JA. (2009) Selective deletion of CD8+ T cells by saporin-coupled MHC class I tetramers. Targeting Trends 10(1).

Penaloza-MacMaster P, Masopust D, Ahmed R (2009) T-cell reconstitution without T-cell immunopathology in two models of T-cell-mediated tissue destruction. Immunology 128:164-171. (Targeting Trends 09q4)


Elson-Schwab L, Garner OB, Schuksz M, Esko JD, Tor Y (2007) Guanidinylated-Neomycin delivers large, bioactive cargo into cells through a heparan sulfate dependent pathway. J Biol Chem 282(18):13585-13591. (Targeting Trends 07q2)

Hess PR, Barnes C, Woolard MD, Johnson MD, Cullen JM, Collins EJ, Frelinger JA (2007) Selective deletion of antigen-specific CD8+ T cells by MHC class I tetramers coupled to the type I ribosome-inactivating protein saporin. Blood 109:3300-3307. (Targeting Trends 07q3)

Yip WL, Weyergang A, Berg K, Tonnesen HH, Selbo PK (2007) Targeted Delivery and Enhanced Cytotoxicity of Cetuximab-Saporin by Photochemical Internalization in EGFR-Positive Cancer Cells. Mol Pharm 4(2):241-251. (Targeting Trends 07q2)


Collins BE, Blixt O, Han S, Duong B, Li H, Nathan JK, Bovin N, Paulson JC (2006) High-affinity ligand probes of CD22 overcome the threshold set by cis ligands to allow for binding, endocytosis, and killing of B cells. J Immunol 177(5):2994-3003. (Targeting Trends 06q4)

Kohls M (2006) Evaluate Potential Targeting Molecules. Nature Methods. (article).

Weyergang A, Selbo PK, Berg K (2006) Photochemically stimulated drug delivery increases the cytotoxicity and specificity of EGF-saporin. J Control Release 111(1-2):165-173. (Targeting Trends 06q2)


Duxbury MS, Ito H, Ashley SW, Whang EE (2004) CEACAM6 as a novel target for indirect type 1 immunotoxin-based therapy in pancreatic adenocarcinoma. Biochem Biophys Res Commun 317(3):837-843. (Targeting Trends 05q3)


Blasius A, Vermi W, Krug A, Facchetti F, Cella M, Colonna M (2004) A cell-surface molecule selectively expressed on murine natural interferon-producing cells that blocks secretion of interferon-alpha. Blood 103(11):4201-4206.


Sheriff ST, Xiao C, Chance WT, Kasckow JW, Balasubramaniam A (2002) Selective lesion of neuropeptide Y (NPY)-receptor neurons in hypothalamus inhibit food intake and reduces body weight in rats. Soc Neurosci Mtg, Orlando FL, Abstract #384.1.