- Home
- Knowledge Base
- References
Enhancement of anti-Robo1 immunotoxin cytotoxicity to head and neck squamous cell carcinoma via photochemical internalization
Komatsu N, Mitsui K, Kusano-Arai O, Iwanari H, Hoshi K, Takato T, Abe T, Hamakubo T (2017) Enhancement of anti-Robo1 immunotoxin cytotoxicity to head and neck squamous cell carcinoma via photochemical internalization. Arch Can Res 5:157-163. doi: 10.21767/2254-6081.100157
Objective: To screen a monoclonal antibody to Robo1, an axon guidance receptor, for its suitability to target various cancers.
Summary: Conventional treatment exhibited an inadequate cytotoxic effect. With the addition of a photosensitizer and LED light illumination, the cytotoxic effect was remarkably improved.
Usage: Saporin-conjugated anti-Robo1 and Saporin-conjugated negative control antibody were prepared by incubating 2 mcl of 1.1 micromolar Streptavidin-ZAP (Biotin Z Internalization Kit) and 2 mcl of 1.1 micromolar biotinylated antibody for 30 min at room temperature.
Related Products: Streptavidin-ZAP (Cat. #IT-27)
Preclinical modeling highlights the therapeutic potential of hematopoietic stem cell gene editing for correction of SCID-X1.
Schiroli G, Ferrari S, Conway A, Jacob A, Capo V, Albano L, Plati T, Castiello M, Sanvito F, Gennery A, Bovolenta C, Palchaudhuri R, Scadden D, Holmes M, 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):eaan0820. doi: 10.1126/scitranslmed.aan0820
Objective: To study potential approaches to gene therapy in mouse models of severe combined immunodeficiency.
Summary: The threshold of IL2RG gene editing can be reached for safe and efficient correction of SCID-X1 established in a preclinical model in human long-term repopulating HSPCs.
Usage: Biotinylated Anti-CD45 was mixed equimolar to Streptavidin-ZAP and administered as a single dose which caused substantial depletion (~70%) of the HSPC compartments and milder depletion of the more mature cell populations.
Related Products: Streptavidin-ZAP (Cat. #IT-27), Anti-CD45.2-SAP (Cat. #IT-91)
Therapeutic Potential of Hematopoietic Stem Cell Gene Editing
A retinoraphe projection regulates serotonergic activity and looming-evoked defensive behaviour.
Huang L, Yuan T, Tan M, Xi Y, Hu Y, Tao Q, Zhao Z, Zheng J, Han Y, Xu F, Luo M, Sollars P, Pu M, Pickard G, So K, Ren C (2017) A retinoraphe projection regulates serotonergic activity and looming-evoked defensive behaviour. Nat Commun 8:14908. doi: 10.1038/ncomms14908 PMID: 28361990
Objective: To investigate how the dorsal raphe nucleus (DRN) and superior colliculus work in concert to extract and translate visual threats into defensive behavioural responses.
Summary: A dedicated population of RGCs signals rapidly approaching visual threats and their input to the DRN controls a serotonergic self-gating mechanism that regulates innate defensive responses.
Usage: Mice received bilateral intraocular injection (2 μg per eye) made between Streptavidin-Saporin and a biotinylated CTB antibody, or Anti-Melanopsin-SAP (2 μg per eye). For detection of melanopsin, retinas were incubated for 3 days at 4 °C with anti-melanopsin (1:600).
Related Products: Streptavidin-ZAP (Cat. #IT-27), Melanopsin-SAP (Cat. #IT-44), Melanopsin Rabbit Polyclonal (Cat. #AB-N38)
Identification of novel macropinocytosing human antibodies by phage display and high-content analysis.
Ha K, Bidlingmaier S, Su Y, Lee N, Liu B (2017) Identification of novel macropinocytosing human antibodies by phage display and high-content analysis. Methods Enzymol 585:91-110. doi: 10.1016/bs.mie.2016.10.004
Objective: To describe a method for identifying antibodies that internalize via macropinocytosis by screening phage-displayed single-chain antibody selection outputs with an automated fluorescent microscopy-based high-content analysis platform.
Summary: Novel phage antibodies are identified by colocalization with macropinocytosis marker, converted into full-length human antibodies, and further characterized with regard to cell binding, pathway of internalization, and intracellular payload delivery.
Usage: Biotinylated IgG is mixed with Streptavidin-ZAP in 1:1 molar ratio to form an immunotoxin that is serially-diluted in a cytotoxicity assay.
Related Products: Streptavidin-ZAP (Cat. #IT-27)
Non-genotoxic conditioning for hematopoietic stem cell transplantation using a hematopoietic-cell-specific internalizing immunotoxin.
Palchaudhuri R, Saez B, Hoggatt J, Schajnovitz A, Sykes D, Tate T, Czechowicz A, Kfoury Y, Ruchika F, Rossi D, Verdine G, Mansour M, Scadden D (2016) Non-genotoxic conditioning for hematopoietic stem cell transplantation using a hematopoietic-cell-specific internalizing immunotoxin. Nat Biotechnol 34:738-745. doi: 10.1038/nbt.3584
Summary: To demonstrate correction of a clinically relevant disease, we employed CD45-SAP in a mouse model of sickle cell anemia and demonstrated our method achieved >90% donor cell chimerism, all mice in three groups (18/18), resulting in complete disease correction (red blood cell counts, hemoglobin levels, hematocrit levels and reticulocyte frequencies were returned to normal). If these pre-clinical results can be successfully translated to the clinic, it would greatly reduce conditioning-related toxicities and expand the use of hematopoietic stem cell transplantation.
Related Products: Streptavidin-ZAP (Cat. #IT-27), Anti-CD45.2-SAP (Cat. #IT-91)
Featured Article: Targeted depletion of hematopoietic stem cells promises safer transplantation
Palchaudhuri R (2016) Featured Article: Targeted depletion of hematopoietic stem cells promises safer transplantation. Targeting Trends 17(3)
Related Products: Streptavidin-ZAP (Cat. #IT-27)
Read the featured article in Targeting Trends.
See Also:
- Palchaudhuri R et al. Non-genotoxic conditioning for hematopoietic stem cell transplantation using a hematopoietic-cell-specific internalizing immunotoxin. Nat Biotechnol 34:738-745, 2016.
- Skulimowska I et al. The biology of hematopoietic stem cells and its clinical implications. FEBS J 16192, 2021.
Method for confirming cytoplaintratumoral anti-HuD immunotoxinsmic delivery of RNA aptamers.
Dickey D, Thomas G, Dassie J, Giangrande P (2016) Method for confirming cytoplaintratumoral anti-HuD immunotoxinsmic delivery of RNA aptamers. (eds. Shum K, Rossi J). In: SiRNA Delivery Methods. Methods in Molecular Biology. 1364:209-217. Humana Press, New York, NY. doi: 10.1007/978-1-4939-3112-5_17
Objective: To describe a functional assay (RIP assay) to confirm cellular uptake and subsequent cytoplasmic release of an RNA aptamer which binds to a cell surface receptor expressed on prostate cancer cells (PSMA).
Summary: This publication details an in vitro functional assay to confirm that the aptamer retains function following conjugation to saporin and describe a cellular assay to measure aptamer-mediated saporin-induced cytotoxicity.
Usage: The folded biotinylated aptamer was mixed at a 1:4 molar ratio of Streptavidin-ZAP, confirmed by agarose gel, a PSMA enzymatic activity (NAALADase) assay performed. FGF-SAP was used as a control.
Related Products: Streptavidin-ZAP (Cat. #IT-27), FGF-SAP (Cat. #IT-38)
Characterization of the first fully macropinocytosing human antibodies human anti-TEM1 scFv in models of solid tumor imaging and immunotoxin-based therapy.
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 macropinocytosing human antibodies human anti-TEM1 scFv in models of solid tumor imaging and immunotoxin-based therapy. Cancer Immunol Immunother 66:367-378.. doi: 10.1007/s00262-016-1937-z
Summary: Tumor endothelial marker 1 (TEM1) has been identified as a novel surface marker upregulated on the blood vessels and stroma in many solid tumors. The authors previous isolated a single-chain variable fragment (scFv) 78 against TEM1 from a yeast display scFv library and evaluated potential applications of scFv78 as a tool for tumor molecular imaging, immunotoxin-based therapy and nanotherapy. MS1 and MS1-hTEM1 cells were treated with site-specifically biotinylated scFv78 conjugated with the Streptavidin-ZAP (Cat. #IT-27) at a molar ratio of 4:1 (scFv78:ZAP) starting from 40 nM serially diluted down to 0.04 nM. The scFv78-saporin immunoconjugate exerted dose-dependent cytotoxicity with high specificity to TEM1-positive cell in vitro. The data indicate that scFv78, the first fully human anti-TEM1 recombinant antibody, recognizes both human and mouse TEM1 and has features advantageous for the development of imaging probes or antibody-toxin conjugates for a large spectrum of human TEM1-positive solid tumors.
Related Products: Streptavidin-ZAP (Cat. #IT-27)
Anti-EFNA4 calicheamicin conjugates effectively target triple-negative breast and ovarian tumor-initiating cells to result in sustained tumor regressions.
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 H, Dylla S (2015) Anti-EFNA4 calicheamicin conjugates effectively target triple-negative breast and ovarian tumor-initiating cells to result in sustained tumor regressions. Clin Cancer Res 21:4165-4173. doi: 10.1158/1078-0432.CCR-15-0695
Summary: Triple-negative breast cancer (TNBC) is characterized by tumors lacking HER2, estrogen receptor, and progesterone receptor. TNBC has proved to be very difficult to treat, in large part because of the absence of consensus targets on the surface of the tumor cells. In this work the authors empirically established a set of surface markers associated with TNBC tumor initiating cells, as produced by patient-derived xenografts. Ephrin-A4 was selected as a therapeutic target, and a cell line transfected with the ephrin-A4 gene was challenged with two versions of biotinylated anti-ephrin-A4 coupled to Streptavidin-ZAP (Cat. #IT-27). Both the mouse monoclonal and the humanized antibodies reach an EC50 of 10 ng/ml, indicating that ephrin-A4 has promise as a therapeutic target for TNBC.
Related Products: Streptavidin-ZAP (Cat. #IT-27)
Intratumoral anti-HuD immunotoxin therapy for small cell lung cancer and neuroblastoma.
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 7:91. doi: 10.1186/s13045-014-0091-3
Summary: HuD protein is a 40-kDa neuronal RNA-binding protein that is expressed in 100% of small cell lung cancer (SCLC) tumor cells. An anti-HuD monoclonal was biotinylated and combined with Streptavidin-ZAP (Cat. #IT-27); this conjugate was tested both in vitro and in vivo. Anti-HuD-SAP eliminated NCI-H69 and Neuro-2a cells at an EC50 of <0.5 μg/ml. 1 mg/kg of the conjugate injected directly into subcutaneous tumors generated in mice resulted in a temporary lack of tumor growth or regression of the tumor. The results demonstrate the potential of HuD as a therapeutic target for SCLC and neuroblastoma.
Related Products: Streptavidin-ZAP (Cat. #IT-27)