What binding and bioactivity data are available for the CD3 epsilon/delta proteins?
Binding and bioactivity data are available across multiple assay platforms, including ELISA, SPR and TR-FRET. For example, CDD-H82W6 binds the reference anti-CD3 antibodies OKT3 (by ELISA and SPR, KD 22.5 nM) and UCHT1 (by ELISA), and supports detection of CD3×BCMA bispecific T-cell engagers in human serum quantitation and TR-FRET assays.
Q
How is the CD3 epsilon/delta heterodimer quality verified?
Heterodimer integrity and purity are confirmed by SEC-MALS (e.g., CDD-H82W6 purity >95% with a molecular weight consistent with the 1:1 heterodimer), and binding activity is validated by ELISA and SPR against reference anti-CD3 antibodies such as OKT3 and UCHT1.
Q
Which CD3 epsilon/delta format should I choose for my assay?
It depends on the detection platform. For streptavidin-based capture assays, choose a biotinylated construct (CDD-H82W6 for human, CDD-C82W6 for cynomolgus). For label-free SPR/BLI, Fc-tagged versions such as CDD-H5255 are suitable. For flow-based binding, fluorescent conjugates such as APC-labeled CDD-HA2H3 or AF647-labeled CDD-HA2W4 are available.
Q
Should I use a CD3 epsilon/delta heterodimer or a CD3 epsilon monomer for binding studies?
A CD3 epsilon monomer is a convenient choice for antibodies that recognize linear or monomeric epitopes. However, some anti-CD3 antibodies recognize conformational epitopes formed when CD3 epsilon associates with CD3 delta; for those clones, the epsilon/delta heterodimer more closely reproduces the native epitope. The choice depends on your antibody's epitope specificity.
Q
What is the CD3 epsilon/delta heterodimer?
The CD3 epsilon/delta heterodimer is a recombinant protein in which the extracellular domains of CD3 epsilon and CD3 delta are co-expressed and assemble into a native-like heterodimer. It reproduces the dimeric epitope recognized by many anti-CD3 antibodies and is widely used to evaluate the CD3-binding arm of T-cell-engaging bispecific antibodies.
Product was of very high quality and worked well for our needs. Served as an effective capture method for our MSD and ELISA assays. Coating with the protein allowed for accurate recovery of antibodies and was consistent between lots and batches ordered.
Logistics speed was as expected, and the product arrived very quickly.
This is one of many biotinylated capture methods we have used and all have functioned as expected and provided consistent results.
This protein was used to immunize mice for antibody development. Proteins were mixed with CFA/IFA and immunized intraperitoneally. Mice were boosted every 14 days for 5 boosts following initial immunization. Serum titers were measured by ELISA following Days 0, 43, and 65. Some signal was detected in mice by Day 43 however by Day 65 all mice had developed an antibody response.
Bought to use it for ELISA. Worked as described. Was impressed by the fast shipping as well as the nice packaging. In addition there was easy to follow protocol accommodated with the product. Each product also came with Lot specific product description sheet ensuring the quality of the product. Worked as described. No doubt tihs five stars product and will will definitely be purchasing again if needed.
This was the first company considered while looking for coating proteins. The protein coating works the way we expect. Doesn't show background. No non-specific binding or signals were observed during ELISA
The purchased protein has been verified to have good biological activity by ELISA assay, and the species can be used for subsequent screening and functional assays of hybridoma monoclonal antibodies.
I had ordered this along with a number of other Acro products, based on good previous experiences with quality and delivery speed, for an immunization campaign and downstream testing.
For us the purity of the product and low endotoxin content were key deciders when choosing to go which product to go for – so it was great to see this provided by Acro.
We had a some delay in receiving part of the order, due to amount ordered, but technical support were great and we were really happy when we received the product and were able to use it, thanks very much Acro!
The product we received last Friday very good condition and shipping pack -20C still freezer. This is a good company supplier for my our company project . Thank you so much from AMSBIO LLC.