Study on the Detection Method of Lipoprotein Phospholipase A2 Based on Nanomicrospheres

Objective: This study is based on the detection of Lp-PLA2 in serum using latex enhanced immunoturbidimetry (PETIA). Reform the existing process by selecting smaller particle size microspheres to increase the upper limit of linearity, selecting microspheres with fewer surface -COOH to ensure low sensitivity, and providing a more accurate and sensitive reagent kit for clinical use. Method: Determine the optimal reaction conditions, including Lp-PLA2 antibody manufacturer, nanoparticle size, antibody concentration, concentration of crosslinking agent [1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDC)], pH value of coupling solution, and concentration of blocking agent. Establish a PETIA method for detecting serum Lp-PLA2. And evaluate its precision, linear range, and recovery rate for performance. Result: The Lp-PLA2 antibody and nanospheres from Huamei Biotechnology were selected as JSR-P0117, with the maximum absorbance range and the best linear relationship at an antibody concentration of 0.2 mg/mL; The optimal reaction conditions for PETIA include a crosslinking agent (EDC) concentration of 95 μg/mL, a blocking agent (10% BSA solution) concentration of 7 μL/mL, and a coupling buffer of boric acid buffer The optimal coupling rate is achieved at a pH value of 8.0. The established PETIA detection sample recovery rate is between 96% - 106%; Within batch coefficient of variation (CV) and between batches CV<2%; The linear range is 10-2100 ng/mL. Conclusion: Comparing to other methods, the PETIA established in this study for detecting Lp-PLA2 in serum is accurate, with a wide linear range, significantly better than the linear range of similar products on the market (20-1000 ng/mL). The experimental results provide laboratory guidance and reference for the development of Lp-PLA2 diagnostic kits.