人载脂蛋白A5单抗制备及临床应用的研究

目的制备抗人载脂蛋白A5（ApoA5）单克隆抗体（McAb）,建立ELISA双抗体夹心法,并进行初步的临床应用。方法利用合成的人ApoA5抗原免疫BALB/c小鼠,制备McAb,并对抗体的效价、特异性、亚类及位点进行了测定,建立了ELISA双抗体夹心法,测定了正常对照组（145例）和冠心病组（68例）血清中ApoA5的含量。结果获得了5个稳定分泌抗体的阳性杂交瘤细胞株,且均为IgG2a型。腹水效价达到1.6×10^5。冠心病组胆固醇（TC）、三酰甘油（TG）、低密度脂蛋白胆固醇（LDL-C）和ApoB水平明显高于对照组（P〈0.05）,而高密度脂蛋白-胆固醇（HDL-C）、ApoA1和ApoA5水平明显低于对照组（P〈0.05）。ApoA5与HDL-C、ApoA1水平呈正相关（r值分别为0.53和0.63,P〈0.05）,与TC、TG、LDL-C、ApoB呈负相关（r分别为-0.69、-0.63、和-0.59,P〈0.05）。结论ApoA5参与脂质代谢,对于心血管疾病的预防和治疗具有一定的临床意义。

Preparation and characterization of monoclonal antibodies against human apolipoprotein A5 and its clinical application

Objective To prepare the monoclonal antibodies against human apolipoprotein A5 （ApoA5）, and to establish a double antibody sandwich enzyme-linked immunoassay for the clinical detection of human ApoA5. Methods The BALB/c mice were immunized with recombinant ApoA5 to prepare the McAb. The antibody titers, specificity, subclass and antigenic determinants of the McAb were identified and measured. The serum ApoA5 levels of 145 healthy people and 68 patients with CHD were detected by ELISA. Results Five hybridoma cell lines secreting McAb against human ApoA5 were obtained, they were highly specific to ApoA5 and all belonged to the subtypes of IgG2a. The ascites titer reached 1.6×10^5. The levels TC,TG,LDL-C and APOB in CHD group were significantly higher than those in control group（ P 〈 0.05 ）, however the levels of HDL-C, ApoA1 and APOA5 in CHD group were significantly lower than those of control group （ P 〈 0.05 ）. Conclusion The APOA5 levels are positively related to HDL-C, ApoA1 levels, however, which are negatively correlated with TC, TG,LDL-C, ApoB levels. To explore the exact relationship between ApoA5 levels and hyperlipidemia may have potential clinical significance in prevention and treatment of cardiovascular diseases.