Phosphocholine bsa pc bsa

Chemiluminescent ELISA of XQ11 binding to indicated antigens was performed in 96 well microtiter plates as previously described (15 ). Red blood cells (RBCs) were obtained from C57BL/6 mice and either used for the experiment as native RBCs or incubated with 0.5% bromelain (Acros, Geel, Belgium) solution for 10 minutes at 37°C. Cells were then washed and stored in PBS at 4°C. For ELISA, 2x10⁵ cells per well were plated. Native LDL and OxLDL were prepared as described (15 ). BSA was from Sigma-Aldrich (St. Louis, MO), and phosphocholine-BSA (PC-BSA) from Biosearch Technologies (Petaluma, CA). XQ11 binding to various antigens was detected using anti-His alkaline phosphatase conjugated antibody from Sigma-Aldrich.

Plasma was obtained from healthy donors after an overnight fast following consent under a protocol approved by the UCSD Human Research Protections Program. LDL was isolated by sequential ultracentrifugation, and modified with malondialdehyde (MDA), malondialdehyde-acetaldehyde-adducts (MAA) or CuSO4 to generate MDA-LDL, MAA-LDL or copper-oxidized LDL (Cu-OxLDL) respectively, as previously described^{31 (link)}. Phosphocholine-BSA (PC-BSA) was from Biosearch Technologies and POVPC-BSA was prepared as described^{32 (link)}. Monoclonal anti-Myc and anti-His alkaline phosphatase (AP) conjugated antibodies were from Sigma.