zap-conjugates

Komatsu N, Kosai A, Kuroda M, Hamakubo T, Abe T (2024) Cetuximab-toxin conjugate and npe6 with light enhanced cytotoxic effects in head and neck squamous cell carcinoma in vitro. Biomedicines 12(5):973. doi: 10.3390/biomedicines12050973 PMID: 38790935

Objective: Combine the use of antibody-directed saporin and a photosensitizer to exert directed and improved cytotoxicity towards carcinoma cells as compared to either by itself.

Summary: Photodynamic therapy uses photosensitizers and irradiation to exert cytotoxic effects on cancer. When combined with biotinylated anti-EGFR conjugated to Strep-ZAP (IT-27), an increased cytotoxicity was hypothesized. The method developed enhanced the cytotoxicity of anti-EGFR-Strep-ZAP by the photodynamic effect in Sa3 and HO-1-u-1 cells, which have moderate levels of EGFR expression.

Usage: Cells were seeded on a 96-well plate and delivered 1.34 pM to 4.2 nM of anti-EGFR or anti-EGFR-Strep-ZAP, and cell viability was measured with formazan.

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

Chen S (2024) Nanocapsule-based prodrugs for targeted treatment of AIDS-associated non-hodgkin lymphoma. Univ California Thesis.

Objective: To propose a novel nanocapsule based platform that encapsulates the native drugs using various monomers and crosslinkers through free radical polymerization.

Summary: This encapsulation technology modifies the surface properties of the encapsulated drugs, enhancing their penetration into deeper tumor tissues and across the blood-brain barrier (BBB). Moreover, it significantly improves the stability of the drugs in vivo, protecting them from rapid degradation or clearance by the immune system. By adjusting the composition of the monomers and crosslinkers, the surface charge, hydrophobicity, and size of the nanocapsules can be finely tuned to maximize their efficacy in reaching and penetrating the target tissues.

Usage: Conjugation of ch128.1Av (anti-TfR1 IgG3-avidin fusion protein) with biotinylated saporin 6 (b-SO6) to eliminate malignant cells, including NHL malignancies. However, safe systemic delivery of ch128.1Av/b-SO6 is limited by its non-specific toxicity to normal cells expressing TfR1.

Related Products: MonoBiotin-ZAP (Cat. #BT-ZAP)

Kitawi R, Ledger S, Kelleher AD, Ahlenstiel CL (2024) Advances in HIV gene therapy. Int J Mol Sci 25(5):2771. doi: 10.3390/ijms25052771 PMID: 38474018

Objective: This review highlights the various stages of ex vivo gene therapy, current research developments that have increased the efficiency and safety of this process, and a comprehensive summary of Human Immunodeficiency Virus (HIV) gene therapy studies, the majority of which have employed the ex vivo approach.

Summary: The long-term or permanent expression of anti-HIV genes and the modification of CD4+ and CD34+ cells to render them resistant to infection or to allow the disruption of the HIV life cycle are important strategies in the quest to achieve a HIV cure.

Usage: Authors referenced CD117 antibody conjugated to saporin via streptavidin (IT-27, IT-83), which enabled >99% depletion of endogenous HSCs in NSG mice and non-human primates.

Related Products: Streptavidin-ZAP (Cat. #IT-27), Anti-CD117-SAP (Cat. #IT-83)

See Also:

• Czechowicz A et al. Selective hematopoietic stem cell ablation using CD117-antibody-drug-conjugates enables safe and effective transplantation with immunity preservation. Nat Commun 10:617, 2019.

Yelamali AR, Chendamarai E, Ritchey JK, Rettig MP, DiPersio JF, Persaud SP (2024) Streptavidin-drug conjugates streamline optimization of antibody-based conditioning for hematopoietic stem cell transplantation. bioRxiv 2024.02.12.579199. doi: 10.1101/2024.02.12.579199 PMID: 38405731

Objective: Use biotinylated CD45.2 antibodies conjugated to Streptavidin to target hematopoietic stem cells (HSCs) for HSC transplantation.

Summary: Hematopoietic stem cell transplantation offers a promising alternative to standard cancer treatments. Using Click Chemistry, different toxic payloads were attached to streptavidin and observed.

Usage: 75 micrograms of CD45.2 delivered to HSCs deriving from B6 mice.

Related Products: Anti-CD45.2-SAP (Cat. #IT-91), Streptavidin-ZAP (Cat. #IT-27)

Zhao Q, Zong H, Zhu P, Su C, Tang W, Chen Z, Jin S (2024) Crosstalk between colorectal CSCs and immune cells in tumorigenesis, and strategies for targeting colorectal CSCs. Exp Hematol Oncol 13(1):6. doi: 10.1186/s40164-024-00474-x PMID: 38254219

Summary: Cancer immunotherapy has become a promising strategy in the treatment of colorectal cancer, and relapse after tumor immunotherapy. Cancer stem cells (CSCs) have the capabilities of self-renewal and differentiation and are also resistant to the traditional therapies of radiotherapy and chemotherapy. The authors review strategies for targeting colorectal CSCs, where one method described uses a biotinylated antibody against EpCAM (clone 3-171) conjugated to saporin via Streptavidin-ZAP (IT-27).

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

See Also:

• Lund K et al. 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):1038-1050, 2014.

Bogen O, Araldi D, Sucher A, Kober K, Ohara PT, Levine JD (2024) Isolectin B4 (IB4)-conjugated streptavidin for the selective knockdown of proteins in IB4-positive (+) nociceptors. Mol Pain 17448069241230419. doi: 10.1177/17448069241230419 PMID: 38246917

Objective: To address the need for selective transfection methods, the authors covalently linked isolectin B4 (IB4) to streptavidin and analyzed whether it could be used to study protein function in IB4(+)-nociceptors

Summary: Rats treated intrathecally with IB4-conjugated streptavidin complexed with biotinylated antisense oligonucleotides for protein kinase C epsilon (PKCε) mRNA werefound to have: (a) less PKCε in dorsal root ganglia (DRG), (b) reduced PKCε expression in IB4(+) but not IB4(–) DRG neurons, and (c) fewer transcripts of the PKCε gene in the DRG. This knockdown in PKCε expression in IB4(+) DRG neurons is sufficient to reverse hyperalgesic priming, a rodent model of chronic pain that is dependent on PKCε in IB4(+)- nociceptors.

Usage: For each intrathecal injection 2.3μL IB4-streptavidin (30 pmol of conjugate with 120 pmol of biotin binding sites), 1.2μL biotinylated ODN (c = 100 pmol/μl) and 16.5μL PBS were mixed (injection volume = 20μL),

Related Products: Streptavidin-ZAP (Cat. #IT-27), IB4-SAP (Cat. #IT-10)

Allred CA, Gormley C, Venugopal I, Li S, McGuire MJ, Brown KC (2023) Tumor-specific intracellular delivery: peptide-guided transport of a catalytic toxin. Commun Biol 6(1):60. doi: 10.1038/s42003-022-04385-7 PMID: 36650239

Objective: The demonstration of a peptide optimized by chemical modifications for tumor specificity to deliver saporin, a catalytic toxin, specifically to cancer cells via both in vitro and in vivo.

Summary: Peptides rival antibodies in affinity and specificity and offer an alternative as cancer-targeting molecules. In comparison to antibodies, peptides have a faster development time and lower production cost. The authors isolated peptide MGS4, derived from a phage-displayed library using a non-small cell lung cancer (NSCLC) cell line as the target. MGS4 was modified to identify the minimal binding domain while also improving affinity and stability. Importantly, the authors provide data showing the peptide delivered saporin both in vitro and in vivo to cancer cells demonstrating anti-tumor efficacy in a mouse model.

Usage: In vitro delivery was performed by reacting biotinylated peptide with Streptavidin-ZAP (Cat. #IT-27) in a 1:1 molar ratio. Cells were treated for 6h at 37C. The drug was removed and replaced with media and after 72 hours, cell viability was measured with CellTiter-GLO. In vivo delivery was performed using biotinylated MGS4 reacted with Streptavidin-ZAP and administered via tail-veil injection (7.5 ug/100 ul) 2x/week for 2.5 weeks for a total of 5 treatments.

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

Heck AJ (2023) Chemical design of a supramolecular protein nanoplatform for therapeutic applications. Univ Johannes Gutenberg Thesis.

Objective: Investigate the potential of using multivalent proteins as an integrative nanoplatform to target protein hallmarks in various diseases by forming supramolecular protein conjugates (SPC).

Summary: The author employs an integrative and adaptable protein-based nanoplatform to combine bioactive entities at the macromolecular level to design multivalent protein-conjugates for versatile therapeutic purposes. Multivalent protein-conjugates, a tetrameric protein with up to four ligand binding sites functions is used as a supramolecular “glue” utilizing avidin-biotin technology. The study investigates the assembly of diverse bioactive components into well organized multifunctional protein-conjugates, ultimately leading to the formation of supramolecular protein conjugates (SPC) that target protein signatures in a variety of diseases.

Usage: Streptavidin-ZAP with Anti-PSMA Antibody

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.

Kirkey DC, Robinson L, Janssens D, Hines MG, Hylkema T, Manselle MK, Ries RE, Peplinski JH, Wallace LK, Otto D, Tarlock K, Henikoff S, Li W, Meshinchi S (2023) CLEC2A is a novel AML-restricted immunotherapeutic target enriched in KMT2A-rearranged acute myeloid leukemia. Blood 142(1):290. doi: 10.1182/blood-2023-188439

Objective: Targeting CLEC2A, a novel immunotherapeutic target, in the treatment of acute myeloid leukemia.

Summary: CLEC2A, a novel immunotherapeutic target expressed highly in acute myeloid leukemia, is a potential target in treating leukemia. By conjugating an antibody targeting this protein to saporin, an ADC for the treatment of leukemia can be created.

Usage: A CLEC2A-antibody was conjugated to Hum-ZAP [IT-22] and treated against CLEC2A+ and CLEC2A- cells and target-specific cytotoxicity was observed at 10nM levels.

Related Products: Hum-ZAP (Cat. #IT-22)

Marquez J (2023) PTGFRN as a target for antibody-drug conjugate (ADC) development in mesothelioma and medulloblastoma. Univ Maryland Baltimore Thesis.

Objective: To investigate the role of Prostaglandin F2 Receptor Negative (PTGFRN) regulator in cancer progression and develop an antibody-drug conjugate (ADC) targeting PTGFRN for the treatment of mesothelioma and pediatric medulloblastoma.

Summary: This dissertation explores the expression and function of PTGFRN in mesothelioma and pediatric medulloblastoma, identifying its association with aggressive cancer phenotypes. The study further develops a potent ADC using a PTGFRN-specific monoclonal antibody conjugated to the cytotoxic compound Duocarmycin, demonstrating significant anti-cancer efficacy in both in vitro and in vivo models .

Usage: Both a custom direct conjugate and Fab-ZAP Mouse (IT-48) were used (up to 10 nM) on transfected HEK-293A cells.

Related Products: Fab-ZAP mouse (Cat. #IT-48), Custom Conjugates