摘要
目的探讨重组人白细胞介素-12(Recombinant human interleukin-12,rhIL-12)与乙型肝炎表面抗原(HBsAg)联合给药对食蟹猴rhIL-12药代动力学的影响。方法以抗rhIL-12单克隆抗体为包被抗体,生物素标记的抗人IL-12单克隆抗体为检测抗体,建立双抗体夹心BAS-ELISA法,测定食蟹猴血清中rhIL-12含量,并对其进行验证。将食蟹猴分为HBsAg(90μg/kg,肌肉注射)与rhIL-12(0.5μg/kg,皮下注射)联用低剂量组和HBsAg(90μg/kg,肌肉注射)与rhIL-12(40μg/kg,皮下注射)联用高剂量组,分别于给药后不同时间采血,分离血清,用建立的BAS-ELISA法测定血清中rhIL-12浓度,采用统计矩方法计算药代动力学参数。结果建立的双抗体夹心BAS-ELISA法的最低定量限为31.25 pg/ml,特异性较强,精密性和准确性良好,检测不同条件处理的样品均保持稳定;联合给药后,食蟹猴血清中rhIL-12的浓度随其剂量的增加而增加,于9 h左右达峰值,高、低剂量组半衰期分别为(21.24±2.03)和(16.05±4.76)h。结论 rhIL-12与HBsAg联合给药对食蟹猴rhIL-12的药代动力学无影响,本研究为IL-12的临床应用奠定了基础。
Objective To investigate the effect of rhIL-12 combined with HBsAg on pharmacokinetics of rhIL-12 in Cynomolgus monkeys.Methods A double antibody sandwich BAS-ELISA method for serum rhIL-12 content in Cynomolgus monkeys was developed by using anti-human IL-12 monoclonal antibody as coating antibody and biotin-labeled anti-human IL-12 monoclonal antibody as detection antibody,and verified.Cynomolgus monkeys were divided into two groups,and injected with 90 μg / kg HBsAg ＋ 0.5 μg / kg rhIL-12 and 90 μg / kg HBsAg ＋ 40 μg / kg rhIL-12 respectively.HBsAg was injected by i.m.route,while rhIL-12 by s.c.route.Serum samples were collected at various time points after injection and determined for rhIL-12 concentrations by the developed BAS-ELISA method,based on which the pharmacokinetic parameters were calculated by statistical moment method.Results The lower limit of quantificaion（LLOQ） of the developed BAS-ELISA method was 31.25 pg / ml.The method showed high specificity,precision and accuracy,by which the determination results of samples treated under various conditions were stable.After injection with HBsAg ＋ rhIL-12,the rhIL-12 concentrations in sera of Cynomolgus monkeys increased with the increasing dosage of rhIL-12 and reached peak values about 9 h after injection.The half-life of rhIL-12 in Cynomlgus monkeys after injection with 90 μg / kg HBsAg ＋ 40 μg / kg rhIL-12 and 90 μg / kg HBsAg ＋ 0.5 μg / kg rhIL-12 were（21.24 ± 2.03） and（16.05 ± 4.76） h respectively.Conclusion The combination of rhIL-12 with HBsAg showed no effect on the pharmacokinetics of rhIL-12 in Cynomolgus monkeys,which laid foundation of clinical application of IL-12.
出处
《中国生物制品学杂志》
CAS
CSCD
2012年第6期718-721,共4页
Chinese Journal of Biologicals
基金
广东省科技计划项目基金(2007B060401053)
广州经济技术开发区科技发展资金(2008Q-P064)
