Targeting Tools: ZAP Kit – How does it work?

Screening large numbers of antibodies for the ability to internalize can be prohibitively expensive in both cost and time. The ZAP Antibody Internalization Kit contains all the components needed for three 96-well plates, or 288 tests. The ability to perform a diagnostic screen that is amenable to high-throughput methods, prior to direct conjugation of those antibodies, is a great cost-benefit in the development of an effective targeted conjugate.

The ZAP Antibody Internalization Kit contains all of the materials needed to screen your antibody. Included, in addition to the selected ZAP products, are controls and XTT developing reagents for the assay. All the user provides are the materials specific to their experiment (the antibody candidate, cells expressing the target, and culture reagents). Recommended protocols for use are detailed in a booklet and on a flash drive provided, and are specific to the particular kit chosen (Whole-ZAP, Fab-ZAP, or FabFc-ZAP). Examples of predicted assay results are also included for comparison; a successful assay provides an EC50 useful in determining if the candidate-antibody should be pursued at the next level.

ATS Customer: “Can you tell me, in a simple way, how I can tell if my antibody internalizes using your kit?”

Brian Russell, Product Manager: “The ZAP technology uses the unique properties of Saporin to verify the binding and internalization of your antibody. Saporin is a ribosome inactivating protein with no binding component, so the only way for Saporin to be toxic to cells is if it is escorted inside the cells via interaction with something else that is internalized (i.e. Your antibody). The ZAP kit measures cell death that is directly and specifically resulting from your antibody being internalized.”

ATS Customer: “How long does it take to get results?”

Brian Russell, Product Manager: “The whole protocol from start to finish takes 5 days. There is an overnight incubation period for cell acclimation and a 72 hour incubation of the cells with your antibody that also gives the Saporin enough time to cause cell death. The hands-on time is approximately 30-60 minutes on each of 3 days during the 5-day protocol.”

ATS Customer: “How many antibodies can I screen at one time?”

Brian Russell, Product Manager: “Each ZAP Kit is designed to test one antibody titrated in sextuplets at 8 concentrations. There are also materials for 1-2 control plates, depending on the kit.”

ATS Customer: “Do you have a kit that will work with other materials — such as peptides?”

Brian Russell, Product Manager: “We do have another product, Biotin-Z Kit (Cat. #KIT-27-Z), that is designed to work with biotinylated materials. If you have a peptide, or other protein that is biotinylated, you can use the Biotin-Z kit as a way to implement ZAP technology when you are working with materials other than antibodies.”

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