United Kingdom (UK)
Recombinant monoclonal antibody to Diethoxyphosphotyrosine. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma DepY.
Immunogen: The diethoxyphosphorylated (OP) peptides cross-linked to four different carrier proteins were used to immunize and boost mice. Mice were immunized with BSA conjugated to 5 peptides diethoxyphosphorylated on tyrosine: PVYSR-OP, LYGLPR-OP, KYAOP, HYRGPA-OP, and EPNVSY-OP. Mice were boosted with OVA conjugated to three peptides diethoxyphosphorylated on tyrosine, RSLYAS-OP, YGGFL-OP, and YPF-OP, and HSA conjugated to PPYRM-OP and QYDVRK-OP. Later on the hybridomas were screened using lysozyme conjugated to RARYEM-OP, GGYR-OP, and KYK-OP.
Specificity: This antibody binds diethoxyphosphotyrosine (chlorpyrifos oxon adduct).
Application Notes: This antibody was characterized in order to determine its binding affinity and binding specificity. The original mouse antibody recognized diethoxyphosphotyrosine independent of the amino acid sequence around the modified tyrosine and independent of the identity of the carrier protein or peptide in an ELISA and western blotting assay. The original mouse IgG2a version of this antibody had an IC50 of 3 × 10−9 M in a competition assay with OP tubulin. Kd values measured by BIAcore and OctetRED96 were 10−8 M for OP-peptides and 1 × 10−12 M for OP-proteins. The limit of detection measured on western blots hybridized with 0.14 μg/mL of depY was 0.025 μg of human albumin conjugated to YGGFL-OP (PMID: 29137457).
Antibody first published in:
Onder et al. Monoclonal Antibody That Recognizes Diethoxyphosphotyrosine-Modified Proteins and Peptides Independent of Surrounding Amino Acids Chem Res Toxicol. 2017 Dec 18; 30(12): 2218–2228. PMID:29137457
Note on publication:
Describes an antibody DepY, which recognizes diethoxyphospho-tyrosine modified proteins and peptides independent of surrounding amino acids. DepY could be useful for identifying new biomarkers of OP exposure. The depY monoclonal antibody is expected to recognize adducts on any protein from any animal.