Preparation of monoclonal antibody to P53 and its clinical application

Objective：The aim of this study was to prepare monoclonal antibody against P53, a kind of tumor suppressor protein,and use the antibody initial y in clinical immunoassay. Methods：Monoclonal antibody was prepared and identified via the classic protocol of monoclonal antibody preparation. Identified monoclonal antibodies were purified by af inity chro-matography. Antibody titer was determined by enzyme linked immunosorbent assay （ELISA）. The specific binding activity of antibody was detected by Western blotting and immunohistochemistry. Results：Three strains of monoclonal antibodies named 1P15, 2P37 and 3P40 were obtained and purified by af inity chromatography. The purity of antibodies was higher than 90%. The titers of antibodies were more than 1：6000. Western blot and immunohistochemistry assay showed that the specific antibody can combine with endogenous P53 protein in the tumor celllines and determine the expression of P53 in tumor tis-sue. Conclusion：Three strains of monoclonal antibodies with high af inity to P53 were successful y established, which can be used for detecting the expression of P53 in tumor cells or tissue.

Microfluidic Immunoprecipitation for Post-Translational Modified Protein Purification

In this paper, we report an antibody functionalized microimmunopreci- pitation （IX IP） method used for enrich lowabundant post-translational modified （PT~ proteins. The device is fabricated by inert, nontoxic and disposable polydimethylsiloxane （PDMS） using a silane-based chemical modification protocol, which yield antibody- terminated PDMS surfaces. In this study, the IX IP device is specifically designed for the purification of carbonylated protein, a representative example here to illustrate the potential applications for any other PTMs, which could be immuno-tagged by specific antibodies. The test model in vitro oxidized bovine serum albumin （BSA） was first derivitized by dinitrophenylhydrazide （DNPH） and then captured by the anti-DNP immobilized on this Ix lP device. The surface functional group mapping was systematically analyzed and validated by fluorescence microscopy. Quantitative study of DNP-derivatized carbonylated protein capture recovery and elution efficiency of the device was also studied. We also envision that this proteome enrichment Ix IP device can be assembled with other lab-on-a-chip components, such as microelectrophoresis or micro-chromatographic devices for follow-up protein analysis. This selective enrichment of modified proteins greatly facilitates the study of low abundant protein biomarkers discovery.

短期新辅助内分泌治疗对前列腺癌神经内分泌分化的影响

目的 探讨短期新辅助内分泌治疗对前列腺癌神经内分泌分化的影响 ,以及神经内分泌分化程度和肿瘤退化程度的关系。方法 采用嗜铬素A(ChA)和 5 羟色胺特异性免疫抗体作为神经内分泌细胞标记物 ,分析 18例前列腺癌患者内分泌治疗前后神经内分泌分化程度的差异。结果治疗前后 ,癌灶内神经内分泌肿瘤细胞数分别为 3.2× 10 5/ μm2 [(0～ 5 .7)× 10 5/ μm2 ]和 2 .3× 10 5/ μm2[(0～ 6 .6 )× 10 5/ μm2 ],差异无显著性 (P >0 .0 5 )。治疗前后神经内分泌肿瘤细胞百分比分别为7.0 % (0 %～ 14 .9% )和 4 .5 % (0 %～ 13.1% ) ,差异无显著性 (P >0 .0 5 )。 6例肿瘤中重度退化者与12例无退化或轻微退化者相比 ,癌灶内神经内分泌肿瘤细胞数差异无显著性 (P >0 .0 5 )。结论 短期新辅助内分泌治疗未导致明显前列腺癌神经内分泌细胞克隆增殖 ,治疗后肿瘤的退化程度和神经内分泌分化程度无明显相关性。