摘要
用人工重组的人前白蛋白多次免疫大耳白兔,获取高效价的前白蛋白抗体。用氯胺T法制备I前白蛋白,再经Sephadex G-25纯化;抗原抗体反应采用平衡一步法,4C培养24 h后经PR试剂分离结合和游离的标记抗原。该法测定范围为12.5-500 ng/ml,最低检出量为20 ng/ml,批内和批间误差分别小于6.4％和8.5％。用该法测得血清前白蛋白人均含量,20-30岁健康人(35例)为(38.49±1.56)μg/ml;70岁以上老人(33例)为(39.24±1.64)ptg/ml;实体肿瘤患者(72例)明显下降为(26.58±2.66)μg/ml。结果表明该法稳定,简便特异,灵敏度适于检出人血清中的前白蛋白水平的变化。
This article is to find a specific and sensitive radioimmunoassay method for prealbumin in human serum. Highly effective antibody of prealbumin was obtained by immunizing rabbits with human recombinant prealbumin many times. The prealbumin antigen was labeled by 125I with chloramines-T methods and purified by the Sephadex G-25 chromatograph column. The reaction between antigen and antibody was carried out by one step balance method and incubated at 4 C for 24 h, then the bond and free antigens were separated by PR reagent. The detection range of this method was from 12. 5 to 500 ng/ ml, the lowest detection level was 20 ng/ml; error within batches and between batches were less than 6. 4% and 8. 5% respectively. The serum prealbumin concentration in health human was (38. 49± 1. 56) μg /ml(n = 35), and in elder (> 70 years) was (39. 24 ± 1. 64) fig/ml(n = 33). The level of prealbumin in patients with solid tumor was significantly lower than that of normal control (26. 58 ± 2. 66) μg/ml(n= 72). It is suggested that this method is stable, simple, specific and highly sensitive,which may be used to test prealbumin in human serum.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2003年第11期1258-1259,共2页
Academic Journal of Second Military Medical University
