用人工重组的白介素6(IL-6)多次免疫兔和豚鼠,获取高效价的IL-6抗体.用氯氨T法制备 125I标记IL-6,经Sephadex G-25纯化,抗原抗体反应采用平衡一步法,4℃温育24 h后经PR试剂分离结合和游离的标记抗原.该法测定范围0.1~3.2 μg/L最低检出量为0.l μg/L,批内和批间误差分别小于6.4%和10%.健康男性115例血清IL-6含量为(0.27±0.13) μg/L.女性101例血清IL-6为(0.26±0.10) μg/L.男女无差异.此外,用该法检测兔失血性休克再灌注损伤后24 h血清IL-6水平明显升高.失血性休克大鼠淋巴液中IL-6水平明显上升,经山莨菪碱(l mg/kg)治疗休克后IL-6又明显下降.内毒素同人牙周纤维细胞共同培养不同时间也促进IL-6释放并明显高于对照水平.
The high effective antibody of interleukin-6 was obtained by immunizing rabbits and guinea pigs with recombinant IL-6 many times. The IL-6 was labeled by 125I with chloramines-T methods and purified by the Sephadex-G25 chromatograph column. The reaction between antigen and antibody was carried out by one step balance method and incubated in 4℃ for 24 hours, then separated bond and free antigen by PR reagent. The detection range of this method was about 0.1~3.2 μg/L, the lowest detection level was 0.1 μg/L, error within batches and between batches was less than 6.4% and 10% respectively. The serum IL-6 concentration in normal male was (0.270±0.13) μg/L (n=115), and in female was (0.260±0.10) μg/L (n=101), there was no difference in male and female group. Otherwise, the level of IL-6 in serum of rabbits was significantly higher than that of self-control at 24 hours after hemorrhagic shock and reperfusion. IL-6 in lymph fluid of rat after hemorrhagic shock was significantly elevated, then it was descended by the treatment of anisodamime (1 mg/kg). The liberation of IL-6 also was promoted and obviously higher than that of the control when fibre cells around tooth were cultured with endotoxin (10 mg/L) at different time in vitro.
颜光涛,郝秀华,王录焕.高灵敏白介素6放射免疫分析的建立及初步应用[J].生物化学与生物物理进展,2000,27(5):541-544
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