摘要
从人体胎盘提纯GST-π,制备兔抗GST-π血清,纯化其IgG,并降解成Fab’片断,通过SMCC与HRP交联成Fab’-HRP,用于夹心ELISA以测定GST-π,灵敏度可达11pg,超过国外的放射免疫法。用此法测定正常人血清GST-π为1.06±0.94ng/ml,原发性肝癌血清较正常高20倍以上。阳性率达90％左右。
GST-π was purified from human placenta and its antisurum was raised in rabbits. The antibody IgG was purified and degraded into Fab' fragment which was conjugated with horseradish peroxidase (HRP) using N-succinimidyl-4-(N-maleimido-methyl) cyclohexane-carboxylate (SMCC) as crosslinking reagent to produce Fab'-HRP conjugate. A sandwich ELISA Was established for the microquantitative determination of GST-π. The sentitivity, was 11pg/ tube, which was far more sensitive than the radioimmunoassay so far reported. Using this method, the serum GST-π of 41 cases of normal adult was found to be 1.06+0.94ng/ml. The upper limit of the normal value was 2.5ng/ml. In 30 cases of primary hepatocarcinoma, the level of serum GST-π was 24.4+17.4ng/ml, which was 23 times higher than the normal average value (p<0.01). The positive rate was 90％. In contrast serum GST-π in 25 cases of chronic hepatitis was determined to be 1.74+1.16ng/ml, which was not significantly different from the normal value (p>0.05). The pseudo-positive rate was 12.0％.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
1990年第5期371-374,共4页
Progress In Biochemistry and Biophysics
关键词
GST
ELISA
肝肿瘤
human placenta type glutathione S-transferase (GST-π), enzyme linked immuno sorbent assay (ELISA), hepatocarcinoma
