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Chemical strategies for antigen-selective targeting of autoreactive B Cells. Chapter 2: Sequential prodrug strategy to target and eliminate ACPA-selective autoreactive B cells.
Lelieveldt L (2019) Chemical strategies for antigen-selective targeting of autoreactive B Cells. Chapter 2: Sequential prodrug strategy to target and eliminate ACPA-selective autoreactive B cells. Radboud Universiteit Nijmegen Nijmegen, Netherlands 45-64. Thesis.
Objective: To develop a method to target and selectively eliminate autoreactive B cells that produce anti-citrullinated proteins antibodies (ACPA) using a sequential antigen prodrug targeting strategy, as a treatment for Rheumatoid Arthritis (RA).
Summary: The study used a synthesized cyclic citrullinated peptide (CCP) antigen suitable for BCR binding and demonstrated that binding by ACPA was impaired upon introduction of a carboxy-pnitrobenzyl (CNBz) caging group at the side chain of the citrulline residue. Enzymatic reduction of the CNBz moiety by nitroreductase fully restored citrulline-selective recognition by both ACPA and ACPA-expressing B cells and showed targeted cell death of CCP-recognizing B cells only. These results mark an important step towards antigen-selective B cell targeting in general and more specifically in RA.
Usage: Streptavidin-ZAP mixed with biotinylated CCP peptides was tested in cytotoxicity assays. The exposure of cells to CCP-SA-ZAP at 1 nM as well as the activated CCP(CNBz) induced death of up to 60% of ACPA-expressing B cells.
Related Products: Streptavidin-ZAP (Cat. #IT-27)
Selective hematopoietic stem cell ablation using CD117-antibody-drug-conjugates enables safe and effective transplantation with immunity preservation.
Czechowicz A, Palchaudhuri R, Scheck A, Hu Y, Hoggatt J, Saez B, Pang WW, Mansour MK, Tate TA, Chan YY, Walck E, Wernig G, Shizuru JA, Winau F, Scadden DT, Rossi DJ (2019) Selective hematopoietic stem cell ablation using CD117-antibody-drug-conjugates enables safe and effective transplantation with immunity preservation. Nat Commun 10:617. doi: 10.1038/s41467-018-08201-x
Objective: To investigate a safe and effective method for hematopoietic stem cell transplantation.
Summary: CD117-ADC selectively targets hematopoietic stem cells yet does not cause clinically significant side-effects.
Usage: The CD117-ADC was prepared by combining biotinylated anti-CD117 (clone 2B8) with Streptavidin–ZAP. A dose of 1.5 mg/kg of CD117-ADC (~12 µg Streptavidin-ZAP) optimally resulted in depletion of >99% of immunophenotypic and functional HSCs.
Related Products: Streptavidin-ZAP (Cat. #IT-27), Anti-CD117-SAP (Cat. #IT-83)
Hematopoietic chimerism and donor-specific skin allograft tolerance after non-genotoxic CD117 antibody-drug-conjugate conditioning in MHC-mismatched allotransplantation.
Li Z, Czechowicz A, Scheck A, Rossi DJ, Murphy PM (2019) Hematopoietic chimerism and donor-specific skin allograft tolerance after non-genotoxic CD117 antibody-drug-conjugate conditioning in MHC-mismatched allotransplantation. Nat Commun 10:616. doi: 10.1038/s41467-018-08202-w
Objective: To develop a conditioning protocol for fully MHC-mismatched bone marrow allotransplantation in mice involving transient immunosuppression and selective depletion of recipient hematopoietic stem cells.
Summary: CD117-ADC conditioning promotes skin allograft tolerance.
Usage: Biotinylated monoclonal antibodies directed against mouse CD117 were coupled to Streptavidin-ZAP. Each mouse was injected with 1.5 mg/kg of ADC in a total volume of 300 mcl PBS.
Related Products: Streptavidin-ZAP (Cat. #IT-27)
Targeting the niche: Depleting haemopoietic stem cells with targeted therapy.
Abadir E, Bryant C, Larsen S, Clark GJ (2019) Targeting the niche: Depleting haemopoietic stem cells with targeted therapy. Bone Marrow Transplant 54:961–968. doi: 10.1038/s41409-019-0445-0
Summary: Anti-mouse CD45 ADC (clone 104-Saporin) Depletes mature lymphoid cells and HSPC, conditioning allows for high level sustained multilineage engraftment of congenic mice. Anti-mouse ADC (CD117-saporin) Combined with T cell depleting agents allowed for significant and durable engraftment in an immunocompetent mouse allo-HSCT model.
Related Products: Streptavidin-ZAP (Cat. #IT-27), Anti-CD45.2-SAP (Cat. #IT-91)
Identification of lineage-specific markers for therapeutic targeting of mast cells.
Plum T (2019) Identification of lineage-specific markers for therapeutic targeting of mast cells. Ruperto-Carola University of Heidelberg, Germany Thesis. doi: 10.11588/heidok.00023555
Usage: Mice were injected i.v. with either 100 µg of 1:1 molar mixture of biotinylated CD63 antibody and Streptavidin-ZAP (60 µg mAb + 40 µg SAP) or with 40 µg SAP alone. All injections were performed in 200 µl PBS.
Related Products: Streptavidin-ZAP (Cat. #IT-27), Saporin (Cat. #PR-01)
Murine HSCs contribute actively to native hematopoiesis but with reduced differentiation capacity upon aging
Säwen P, Eldeeb M, Erlandsson E, Kristiansen TA, Laterza C, Kokaia Z, Karlsson G, Yuan J, Soneji S, Mandal PK, Rossi DJ, Bryder D (2018) Murine HSCs contribute actively to native hematopoiesis but with reduced differentiation capacity upon aging. Elife 7:e41258. doi: 10.7554/elife.41258 PMID: 30561324
Related Products: Streptavidin-ZAP (Cat. #IT-27)
Human anti-NKp46 antibody for studies of NKp46-dependent NK cell function and its applications for type 1 diabetes and cancer research.
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. (2019) Human anti-NKp46 antibody for studies of NKp46-dependent NK cell function and its applications for type 1 diabetes and cancer research. Eur J Immunol 49(2):228-241. doi: 10.1002/eji.201847611
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.
Usage: Conjugation of the antibodies to Saporin, treatment of cells, and cell viability assay Biotin-Z Kit instructions.
Related Products: Streptavidin-ZAP (Cat. #IT-27)
Sequential prodrug strategy to target and eliminate ACPA-selective autoreactive B cells.
Lelieveldt LPWM, Kristyanto H, Pruijn GJM, Scherer HU, Toes REM, Bonger KM (2018) Sequential prodrug strategy to target and eliminate ACPA-selective autoreactive B cells. Mol Pharm 15(12):5565-5573. doi: 10.1021/acs.molpharmaceut.8b00741
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.
Usage: Biotinylated CCP1, CArgP1, and CCP1(CNBz) were conjugated with Streptavidin-ZAP in a 4:1 ratio to make peptide-drug conjugates.
Related Products: Streptavidin-ZAP (Cat. #IT-27)
Antibody drug conjugates targeted to CD45 or CD117 enable allogeneic hematopoietic stem cell transplantation in animal models
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. Blood 132:3324. doi: 10.1182/blood-2018-99-119432
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-SAP and CD117-SAP were more effective at conditioning versus 2Gy TBI or pretransplant Cytoxan.
Usage: 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).
Related Products: Streptavidin-ZAP (Cat. #IT-27)
Screening targeting agents and their cell surface biomarkers for high specificity and rapid internalization via cell death and fluorescence
Ancheta L, Bouajram R, Lappi DA (2018) Screening targeting agents and their cell surface biomarkers for high specificity and rapid internalization via cell death and fluorescence. Neuroscience 2018 Abstracts 128.20 / M17. Society for Neuroscience, San Diego, CA.
Summary: Some of the most recent successes in the treatment of cancers or research into passive immunotherapies for neurodegenerative diseases, employ the use of antibodies. These treatments utilize antibodies that either: 1) interfere with cell surface proteins responsible for tumor cell proliferation, 2) act as immune checkpoint inhibitors, or 3) are re-engineered to allow transport of other molecules across the blood-brain barrier (BBB). There are a growing number of antibody and small molecule therapeutic candidates and this demands a quick and efficient technique to screen for biomarkers that internalize effectively upon binding. The method described provides for the efficient determination of internalization of cell surface biomarkers upon binding of antibodies or peptides. This one-step, robust method uses a targeting agent combined with both a fluorescent reporter and a cytotoxic payload. The construct that makes this method effective was formed by cross-linking a fluorescent reporter, in this case fluorescein (FITC) and streptavidin to the ribosome-inactivating protein, Saporin. The conjugate used in screening potential therapeutics is a mixture of a biotinylated targeting agent mixed in a 1:1 molar ratio with FITC-labeled Streptavidinylated-Saporin. The method provides a definitive assay readout: fluorescence within 1 hour and cell death in 72 hours. This method is designed for rapid screening, in a quick and reproducible manner, for specificity and internalization in various cell types to explore suitability of candidates as therapeutics.
Related Products: Streptavidin-ZAP (Cat. #IT-27), FITC-Streptavidin-ZAP (Cat. #IT-85)
See Also:
- Tan HL et al. Conservation of oncofetal antigens on human embryonic stem cells enables discovery of monoclonal antibodies against cancer. Sci Rep 8:11608, 2018.
- Yuan X et al. 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, 2017.
- Forsyth PA et al. p75 neurotrophin receptor cleavage by α- and γ-secretases is required for neurotrophin-mediated proliferation of brain tumor-initiating cells. J Biol Chem 289(12):8067-8085, 2014.
- Thakkar JP et al. Epidemiologic and molecular prognostic review of glioblastoma. Cancer Epidemiol Biomarkers Prev 23(10):1985-1996, 2014.
- Kohls M Evaluate Potential Targeting Molecules. Nature Methods , 2006.