【摘要】 本文建立了肿瘤相关胰蛋白酶原 - 2 (TAT - 2 )IRMA ,并初步应用于临床 ,以探讨其对肿瘤早期诊断的可能性。以抗TAT - 2McAbT2C和T2L分别作为捕捉抗体和标记抗体。在 pH9.6的碳酸缓冲液中 ,将McAbT2C包被小珠 ,成为固相抗体 ;用氯胺T方法将McAbT2L制成1 2 5I -标记抗体 ;并将两株McAb与不同量的TAT - 2标准品 ,在pH7.5的磷酸缓冲液中保温反应 ,制作TAT - 2标准曲线。检测肿瘤患者及正常人血清或尿液TAT - 2含量 ,并作对比。结果表明 ,测定的TAT - 2标准曲线 (N =1 0 ) :Bmax/T =(2 2± 3.2 ) % ,Bmax/B0=6 89.2± 35 .6倍 ,非特异结合率小于 0 .5 % ,检测下限在 0 .5ng/mL ;批内CV为 (4 .3± 0 .2 6 ) % ,批间CV为(1 0 .6± 1 .2 ) % ;平均回收率为 (98.5± 3.1 ) %和 (97.9± 2 .3) %。正常人血清 (87份 )和尿 (2 0份 )TAT - 2浓度为 7.5 8± 5 .4 6ng/mL和 3.1 1± 1 .6 2ng/mL ;胃癌患者 (4 5例 )的血清TAT - 2浓度 6 3.2± 6 2 .9ng/mL ,胰腺癌和胆管癌患者 (1 2例 )和其它各种转移癌患者 (1 6例 )尿TAT - 2浓度为 2 36 .6± 1 85 .3ng/mL ,1 0 4 .9± 1 71 .2ng/mL ;3组癌症患者血清、尿TAT - 2浓度与正常人相比 ,差别非常显著 (P值均小于 0 .0 0 1 )。因此 ,TAT - 2有希望成为一项肿瘤辅助诊断指?更多还原

【Abstract】 To develop the immunoradiometric assay(IRMA) for tumor-associated trypsinogen-2(TAT-2), and trying to apply it in cancer diagnosis, the anti-TAT-2 McAb T2C and T2L were used as catching and labeling antibody, respectively.The polystyrene beads were coated with McAb T2C to form solid phase in carbonate buffer of pH9.6; the McAb T2L was iodinated with 125 I by chloramine-T. The different amounts of standard TAT-2 were reacted with solid phase McAb and 125 I-labeled McAb to establish standard curve.The serum or urine TAT-2 levels of cancer patients and normal individuals were detected and compared with each other.Results showed that the B max /T was (22±3.2)%, B max /B 0 ratio was (689.2±35.6)%, the non-specific binding rate was below 0.5%, and the low detected limit was 0.5ng/mL.The within-assay CV was (4.3±0.26)%, the between-assays CV was (10.6±1.2)%.The recoveries of detection of serum and urine TAT-2 were (98.5±3.1)% and (97.9±2.3)%, respectively.The TAT-2 value in mormal individuals in sera (n=87) was 7.58±5.46ng/mL, and in urine(n=20) was 3.11±1.62ng/mL.The TAT-2 value in patients with gastric cancer in sera was 63.12±62.9ng/mL.The TAT-2 value in patients with pancreatic and bile duct cancers(n=12)in urine was 263.6±185.3ng/mL, and TAT-2 in patients with other metastatic cancers(n=16)in urine was 104.9±171.2ng/mL.The differences of TAT-2 levels between cancer patients and normal individuals were very significant( P <0.001 for all three groups).Therefore, the development of TAT-2 IRMA could provide a good indicator which should be very helpful in cancer diagnosos.更多还原