Kristen Hartman – Page 17 – Advanced Targeting Systems

Upregulation of p75NTR by histone deacetylase inhibitors sensitizes human neuroblastoma cells to targeted immunotoxin-induced apoptosis

Dedoni S, Olianas A, Manconi B, Collu M, Tuveri B, Vincis ME, Olianas MC, Onali P (2022) Upregulation of p75NTR by histone deacetylase inhibitors sensitizes human neuroblastoma cells to targeted immunotoxin-induced apoptosis. Int J Mol Sci 23(7):3849. doi: 10.3390/ijms23073849 PMID: 35409209 Summary: Histone deacetylase (HDAC) inhibitors have been useful chemotherapy agents in the treatment of […]

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Dosage of Fab-ZAP for antibody concentration

Kristen Hartman

Q: Is the dosage of Fab-ZAP always enough for any level of antibody concentration? A: The 4.5 nM dosage of Fab-ZAP is the recommended concentration. We do not typically see unspecific killing at 4.5 nM on most cell lines. If the concentration of Fab-ZAP is increased, it may undergo bulk-phase endocytosis and kill cells indiscriminately.

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Recommended ratio between Fab-ZAP dosage and antibody concentration

Kristen Hartman

Q: Is there a recommended ratio between Fab-ZAP dosage and antibody concentration? A: A recommended good starting point is 4.5 nM of Fab-ZAP, with a titration of the antibody starting at a concentration of 10 nM. Related Products: ZAP Conjugates

Fab-ZAP number of replications

Kristen Hartman

Q: Each concentration is suggested to perform 6 replications, can it be adjusted more or less in practice? A: Yes, the assay design is meant to be a thorough approach but can be adjusted by the user. We recommend 6 replications based on our 96-well plate template design. The concentration of Fab-ZAP is 4.5 nM

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Detecting the targeted antibody in supernatant

Kristen Hartman

Q: Can Fab-ZAP detect the targeted antibody still in supernatant? A: As long as there is nothing in the supernatant inhibiting the reactivity of Fab-ZAP, it should work. We do not typically recommend this, but in theory it should be possible. I would be cautious of this approach based off of the presumed lack of

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DMSO% for peptide and Streptavidin-ZAP

Kristen Hartman

Q: Instead of performing the reaction between our biotinylated peptide and Streptavidin-ZAP at the initially provided concentration of Strep-ZAP (20 µM), is it OK if the reaction is done at a 10-fold more dilute concentration? This request is to ensure we don’t have any solubility problems with our very tricky lipophilic peptide. Our protocol would

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Urokinase-type plasminogen activator receptor (uPAR) as a therapeutic target in cancer

Kristen Hartman

Zhai BT, Tian H, Sun J, Zou JB, Zhang XF, Cheng JX, Shi YJ, Fan Y, Guo DY (2022) Urokinase-type plasminogen activator receptor (uPAR) as a therapeutic target in cancer. J Translational Medicine 20:135. Summary: The authors describe how a conjugate of Saporin and ATF (Urokinase-type plasminogen activator receptor (uPAR) as a therapeutic target in

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Immunological barriers to haematopoietic stem cell gene therapy

Kristen Hartman

Charlesworth CT, Hsu I, Wilkinson AC, Nakauchi H (2022) Immunological barriers to haematopoietic stem cell gene therapy. Nat Rev Immunol 1-15. doi: 10.1038/s41577-022-00698-0 Objective: This review article attempts to encourage more research to address the immunological barriers to haematopoietic stem cell based gene therapies. Summary: The authors lay out the history and clinical trials of

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A CD45-targeted antibody-drug conjugate successfully conditions for allogeneic hematopoietic stem cell transplantation in mice

Kristen Hartman

Saha A, Hyzy S, Lamothe T, Hammond K, Clark N, Lanieri L, Bhattarai P, Palchaudhuri R, Gillard GO, Proctor J, Riddle MJ, Panoskaltsis-Mortari A, MacMillan ML, Wagner JE, Kiem HP, Olson LM, Blazar BR (2022) A CD45-targeted antibody-drug conjugate successfully conditions for allogeneic hematopoietic stem cell transplantation in mice. Blood 139(11):1743-1759. doi: 10.1182/blood.2021012366 PMID: 34986233

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ZAP Internalization Kit Concentrations

Kristen Hartman

Q: We have your ZAP internalization kit and I have a question regarding the concentrations used in the cytotoxicity assay. The Hum-ZAP used in the assay (mentioned in the PDF protocol) is 4.5 nM and the target agent was 10 nM to 1 fM. Is there a stoichiometric relation between Hum-ZAP and the target agent

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Author name: Kristen Hartman