Development and application of monoclonal antibodies anti-human lipoprotein-associated phospholipase A2.
- Author:
Wen XU
1
;
Zhaoxin LU
1
;
Lin CAO
1
Author Information
- Publication Type:Journal Article
- Keywords: atherosclerosis; eukaryotic expression; immunochromatography; lipoprotein-associated phospholipase A2; monoclonal antibody
- MeSH: 1-Alkyl-2-acetylglycerophosphocholine Esterase; metabolism; Animals; Antibodies, Monoclonal; Enzyme-Linked Immunosorbent Assay; Humans; Mice; Mice, Inbred BALB C
- From: Chinese Journal of Biotechnology 2019;35(3):482-491
- CountryChina
- Language:Chinese
- Abstract: The aim of this study is to prepare monoclonal anti-human Lp-PLA2 antibodies, and establish a rapid and accurate immunochromatographic Lp-PLA2 assay used in community medical institution. The gene sequence of human Lp-PLA2 was obtained from NCBI to construct the expression plasmid. Lp-PLA2 protein expressed in CHO-K1 cells was used to immune BALB/c mice. The monoclonal antibodies were produced in mouse ascites after hybridoma cells screening. Antibodies were evaluated by SDS-PAGE, ELISA and other methods. The Lp-PLA2 test strip was prepared based on sandwich method and evaluated with the portable detection instrument. The affinity of the paired antibodies, PLA1 and PLA5, both reached 1×10⁻⁸. The antibody subclass was IgG1. Both antibodies recognized the Lp-PLA2 protein in the blood specifically. The Lp-PLA2 test strip was prepared based on sandwich method, with linear range of 20-2000 ng/mL. The Lp-PLA2 test strip correlated well with the diaDexus ELISA test kit. In conclusion, the paired antibodies were successfully prepared with high affinity and specificity. The immunochromatographic test of Lp-PLA2 provided a fast and accurate method to detect the concentration of Lp-PLA2 in blood sample for clinical use in the community medical institution and could contribute to the management of cardiovascular diseases.
