zap-conjugates

Su Y, Liu Y, Behrens CR, Bidlingmaier S, Lee NK, Aggarwal R, Sherbenou DW, Burlingame AL, Hann BC, Simko JP, Premasekharan G, Paris PL, Shuman MA, Seo Y, Small EJ, Liu B (2018) Targeting CD46 for both adenocarcinoma and neuroendocrine prostate cancer. JCI Insight 3(17):e121497. doi: 10.1172/jci.insight.121497 PMID: 30185663

Objective: To investigate the suitability of a CD46 antibody as a metastatic castration-resistant prostate cancer (mCRPC) anti-tumor agent.

Summary: CD46 is an excellent candidate for antibody-based therapy development, which has potential to be applicable to both adenocarcinoma and neuroendocrine types of mCRPC that are resistant to current treatment.

Usage: Biotinylated UA20 IgG and Streptavidin-ZAP were mixed at a molar ratio of 1:1. The conjugate was used in cytotoxicity assays and shown to specifically kill the mCRPC cells.

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

Tan HL, Yong C, Tan BZ, Fong WJ, Padmanabhan J, Chin A, Ding V, Lau A, Zheng L, Bi X, Yang Y, Choo A (2018) Conservation of oncofetal antigens on human embryonic stem cells enables discovery of monoclonal antibodies against cancer. Sci Rep 8:11608. doi: 10.1038/s41598-018-30070-z

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.

Usage: in 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. invivo – 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.

Related Products: Hum-ZAP (Cat. #IT-22), Streptavidin-ZAP (Cat. #IT-27), Saporin (Cat. #PR-01)

Hoffmann RM, Crescioli S, Thurston DE, Karagiannis SN (2018) Development and evaluation of T-Zap: a novel antibody-drug conjugate for the treatment of Her2 positive breast cancer. Cancer Res 78:LB-001. doi: 10.1158/1538-7445.AM2018-LB-001 PMID: 909090

Objective: Develop and Evaluate a novel ADC (T-Zap) for breast cancer.

Summary: Binding to target cells of T-Zap was confirmed. Comparison of T-Zap efficacy in breast cancer cell lines with and without resistance against trastuzumab showed a trend for higher efficacy of cell killing by T-Zap in trastuzumab resistant cells compared to T-DM1. Toxicity assays revealed no impact of T-Zap on cell viability in immune cells.

Usage: T-ZAP was made using Biotinylated monoclonal antibody trastuzumab mixed with Streptavidin-ZAP.

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

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)

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)

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)