zap-conjugates

Harvey BP, Cohen-Solal J, Kaymakcalan Z (2018) SAT0058 Adalimumab: TNF complexes are cleared more efficiently by human osteoclasts than those with etanercept through FCG-receptor binding and internalisation. Ann Rheum Dis 77:893. EULAR 2018, Amsterdam, The Netherlands doi: 10.1136/annrheumdis-2018-eular.3804

Objective: To determine whether Fc-gamma receptor (FcgR)-mediated internalization of the biologic:TNF complexes is a contributing mechanism responsible for the difference in effectiveness between ADA and ETN in preventing TNF- enhanced OCgenesis.

Summary: Human osteoclast  (OC) precursors can bind and internalise ADA:TNF complexes more efficiently than ETN:TNF complexes. In addition, this process is partially mediated through FcgRII.

Usage: FcgR-mediated nternalization was assessed by monitoring a reduction in OC survival in response to preformed bio- logic: TNF complexes (25:1 ratio) bound with FabFC-ZAP human ± FcgR blocking antibodies.

Related Products: FabFc-ZAP human (Cat. #IT-65)

Sakamoto A, Kato K, Hasegawa T, Ikeda S (2018) An agonistic antibody to EPHA2 exhibits antitumor effects on human melanoma cells. Anticancer Res 38:3273-3282. doi: 10.21873/anticanres.12592

Objective: Investigate the therapeutic potential of antibody to EPHA2 against melanoma in vitro.

Summary: Observations indicate a promising role for EPHA2 as a target in antibody treatments for melanoma, and demonstrate the potential therapeutic effects of an agonistic antibody to EPHA2.

Usage: A375 cells were plated into a flat-bottom, 96-well plate (2,000 cells per well) and incubated for 4 days at 37˚C. Cell suspension included different concentrations of Mab-ZAP, along with either anti-EPHA2 mAb (SHM16, SHM17, or SHM20 at 2 μg/ml final concentration), or a control IgG1 mAb (2 μg/ml final concentration).

Related Products: Mab-ZAP (Cat. #IT-04)

Sadhukhan R, Brown N, Ouellette D, Banach D, Filoti DI, Winarta D, Raghavendra R, Sousa S, Darcy A, Alessandri L, Ivanov A, Bose S, Eaton L, Preston G, Freeman J, Correia I (2018) Engineering elastic properties into an anti-TNFα monoclonal antibody. Cogent Biol 4(1):1469387. doi: 10.1080/23312025.2018.1469387

Objective: To engineer elastic properties into a TNFalpha antibody.

Summary: The results presented in this report with an anti-TNFα ELP mAb are a foundation for building on a new generation of fusion ELP mAbs, or other formats, that are stable, active, responsive to cues in local environment, and, with the FcRn mutation, cleared rapidly from circulation. More detailed studies are warranted to identify the appropriate ELP sequences for IA delivery, calculate residence time in the IA space, and demonstrate pharmacodynamics effect of the ELP-fusion protein.

Usage: Fab-ZAP human was mixed with anti-TNFα-ELP fusion monoclonal.

Related Products: Fab-ZAP human (Cat. #IT-51)

Wüstemann T, Haberkorn U, Babich J, Mier W (2019) Targeting prostate cancer: Prostate-specific membrane antigen based diagnosis and therapy. Med Res Rev 39(1):40-69. doi: 10.1002/med.21508 PMID: 29771460

Summary: Conjugation to the antibody was achieved by reacting the biotinylated humanized antibody to prostate-specific membrane antigen (PMSA) with Streptavidin-ZAP. Binding potency of the conjugate was comparable to that of the naked antibody and in vivo experiments proved potent for selective tumor growth inhibition in mice bearing LNCaP tumors.

Related Products: Streptavidin-ZAP (Cat. #IT-27)

See Also:

• Kuroda K et al. Saporin toxin-conjugated monoclonal antibody targeting prostate-specific membrane antigen has potent anticancer activity. Prostate 70(12):1286-1294, 2010.

Eng MS, Kaur J, Prasmickaite L, Engesaeter BO, Weyergang A, Skarpen E, Berg K, Rosenblum MG, Maelandsmo GM, Hogset A, Ferrone S, Selbo PK (2018) Enhanced targeting of triple-negative breast carcinoma and malignant melanoma by photochemical internalization of CSPG4-targeting immunotoxins. Photochem Photobiol Sci 17:539-551. doi: 10.1039/C7PP00358G

Summary: The combination of the drug delivery technology PCI and CSPG4-targeting immunotoxins is an efficient, specific and light-controlled strategy for the elimination of aggressive cells of TNBC and malignant melanoma origin. This study lays the foundation for further preclinical evaluation of PCI in combination with CSPG4-targeting.

Usage: To obtain the immunotoxin 225.28-saporin, Streptavidin-Saporin (Cat. #IT-27; Streptavidin-ZAP), with an average of 2.5 molecules of saporin per molecule of streptavidin, was combined with biotinylated 225.28, a CSPG4-specific mouse mAb, IgG2a.

Related Products: Streptavidin-ZAP (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 (2018) Characterization of the first fully human anti-TEM1 scFv in models of solid tumor imaging and immunotoxin-based therapy. Cancer Immunol Immunother 67:329-339. doi: 10.1007/s00262-017-2101-0 PMID: 29313073

Objective: ScFv78 was conjugated with the ribosome-inactivating protein saporin (Streptavidin-ZAP) to evaluate whether scFv78 may be used as a vehicle for theTEM1-targeted delivery of toxins.

Summary: Site-specific, biotinylated scFv78 was conjugated with streptavidin-labeled saporin (Streptavidin-ZAP; Cat. #IT-27) by incubation at room temperature for 1h at a molar ratio of 4:1 (scFv78:ZAP).

Usage: Mouse endothelial cells (MS1) and MS1 cells transduced to express full-length human TEM1 (MS1-TEM1) were cultured in 96-well plates to 30% confluence and then incubated for 96h in the presence of 10-fold serially diluted Streptavidin-ZAP, scFv78, or scFv78-ZAP starting from 40nM down to 0.04nM. The data indicate that scFv78, the first fully human anti-TEM1 recombinant antibody, recognizes both human and mouse TEM1 and 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.

Related Products: Streptavidin-ZAP (Cat. #IT-27)

Kusano-Arai O, Iwanari H, Kudo S, Kikuchi C, Yui A, Akiba H, Matsusaka K, Kaneda A, Fukayama M, Tsumoto K, Hamakubo T (2018) Synergistic cytotoxic effect on gastric cancer cells of an immunotoxin cocktail in which antibodies recognize different epitopes on CDH17. Monoclon Antib Immunodiagn Immunother 37:1-11. doi: 10.1089/mab.2017.0043

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.

Usage: The authors applied a monoclonal antibody (mAb) cocktail for one target protein with multiple epitopes. They generated anti-CDH17 mAbs recognizing different epitopes on CDH17 (Cadherin-17). CDH17 is expressed in gastric cancer, hepatocellular carcinoma, colorectal cancer, and pancreatic cancer and has limited distribution in normal tissues. For preparation of 3 immunotoxins, Streptavidin-ZAP was mixed with biotinylated mAbs in equimolar concentrations for 30 minutes at room temperature. The study provides data to demonstrate that the cocktail of different epitope-recognizing immunotoxins has synergistic cytotoxic effects on CDH17-expressing cells.

Related Products: Streptavidin-ZAP (Cat. #IT-27)

Cua S, Tan HL, Fong WJ, Chin A, Lau A, Ding V, Song Z, Yang Y, Choo A (2018) Targeting of embryonic annexin A2 expressed on ovarian and breast cancer by the novel monoclonal antibody 2448. Oncotarget 9:13206-13221. doi: 10.18632/oncotarget.24152

Objective: To develop mAbs to potentially target oncofetal antigens and be repurposed for antibody or antibody drug conjugate (ADC) therapy.

Summary: The novel IgG1, 2448, was shown to target a unique glycosylated surface epitope on ANXA2. As a possible therapeutic candidate for ovarian and breast cancer, 2448 demonstrated anti-tumor activity via two independent mechanisms of action.

Usage: Cells were seeded in 96-well plates at 1000 or 2000 cells/well. Primary antibody, 2448 or ch2448 (10 μg/mL) was pre-mixed with appropriate secondary saporin conjugate, Mab-ZAP or Hum-ZAP.  The most significant decreases in cell viability (20% to 60%) were observed against the epithelial IGROV1 and MCF7 cell lines.  ATS created a Custom ADC by direct conjugation of saporin to ch2448 (ch2448-SAP).  As a control, an isotype chimeric IgG was also conjugated to saporin (IgG-SAP). Compared to using secondary saporin conjugates, ch2448-SAP induced and increase of  20–30% cytotoxicity.)

Related Products: Mab-ZAP (Cat. #IT-04), Hum-ZAP (Cat. #IT-22), Custom Conjugates

Su Y, Bidlingmaier S, Lee NK, Liu B (2018) Combine phage antibody display library selection on patient tissue specimens with laser capture microdissection to identify novel human antibodies targeting clinically relevant tumor antigens. (eds. Hust M, Lim T). In: Phage Display. Methods in Molecular Biology. 1701:331-347. Humana Press, New York, NY. doi: 10.1007/978-1-4939-7447-4_18

Objective: To develop a technology that allows selection of phage antibody display libraries on tumor cells in situ residing in their natural tissue microenvironment.

Summary: Intracellular delivery of Immunotoxin was determined as follows: Immunotoxin was prepared by mixing biotinylated scFv with Streptavidin-ZAP (Cat. #IT-27) at a molar ratio of 1:1 and incubated on ice for 30 min. 50 μl of serially diluted immunotoxin was added to each well and incubated for 96 h at 37°C in 5% CO2. Cell growth medium were carefully removed from each well.

Usage: 100 μl of diluted CCK-8 was added to each well in the 96-well plates and incubated for 1–4 h at 37°C in 5% CO2. The absorbance was measured at 450 nm using a microtiter plate reader and the EC50 value determined using GraphPad Prism.

Related Products: Streptavidin-ZAP (Cat. #IT-27)

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)