Abstract
Binding capacities of immunoassay reagents for remnant-like particles, mixed gel suspension of anti-apo B-100 and apo A-I monoclonal antibodies coupled to Sepharose 4B, to LDL and HDL were examined by HPLC method. The resolution and sensitivity for detection of cholesterol (C) in serum lipoproteins have been improved using a new special column (TSK gel Lipopropak, 7.5 mm I. D., 600 mm length / TOSOH Co. Tokyo) as well as a new eluent (TSKeluent LP-1). Within-run precision of this method by applying 100 μl of highly diluted serum (× 1,001) was excellent both for LDL-C and HDL-C (CV value: 2.49% and 2.48 %). About 98% of LDL and HDL of serum specimens containing less than 210 mg/dl of LDL-C, and 100 mg/ dl of HDL-C was bound to the immunoassay reagent gel. Therefore, in the routine assay technique, in which 5 μl of serum was added to 300μl of mixed gel suspension and the reaction mixture was gently shaken for 60 min at room temperature, maximun binding capacities of the reagent gel for LDL-C and HDL-C were suggested to be 210mg/dl and 100mg/dl, respectively.
