Preparation of apolipoprotein M monoantibody and its preliminary application / 中华检验医学杂志

ABSTRACT

Objective To prepare anti-apoM monoantibodies with high affinity and high purity, and investigate apoM distribution among human tissues and different groups of people. Methods BALB/c mice were injected intracutaneously with recombinant apoM. After cytomixis, screening and cloning, we established a hybridoma, which grew well and steadily secreted antibodies. The ascites were acquired by injecting BALB/c mice intraperitoneally and anti-apoM monoantibodies were gained using standard techniques. We detected apoM levels in healthy individuals and the patients with coronary heart disease including stable angina (SA) group and acute coronary syndrome (ACS) group using the anti-apoM monoantibodies. ResultsThree anti-apoM monoantibodies were collected and confirmed after subtype identification and block test. The apoM protein were detected in some human cells and human tissues by these three monoantibodies. The concentration of apoM was (11.02 ±1.96) ×10 -3 g/L, (10. 76± 1.32) ×10-3 g/L, (12. 83 ± 2. 28) × 10-3 g/L in SA, ACS and control group respectively. There was significant difference within the three groups (F = 11. 544, P ＜ 0. 05). Comparing apoM concentrations among control group and coronary heart disease groups, it showed that the levels of apoM were lower in coronary heart disease groups than in control group(t =2. 962 and 3. 967,P ＜0. 05). There was no significant difference between two coronary heart disease groups (t = 1. 033, P ＞ 0. 05). Conclusion Anti-apoM monoantibodies are successfully raised and could combine with apoM in human cells and tissues. This lays the foundation for the apoM study in apolipoprotein metabolism.