纯化

The AmkA, AmrA and AmrB had been purified with the His-Tag chromatography column.

其中，amrA，amkA和amrB基因的表达产物已经His-Tag亲和柱进行了纯化。

The ELISA titre was 1:2000.By cell fusion, 46 hybridoma cell lines were screened,and 10 lines were cloned with limited dilution method.16 lines secreting anti-bFGF monoclonal antibody were been developed, and 2 lines targeted fusion protein. Sensitive ELISA and dot-ELISA for bFGF was developed with this mAb. The detection limit of them were 0.1 ng/well and 0.5 ng/well. The expression level of anti-bFGF mAb by different rebuilt engineering cells were identified by western blot and to direct rebuild recombiment engineering cell. The dose and character of anti-bFGF mAb inhibiting bFGF biology activity were searched by 3T3 cell line. Searching 20 tissue of liver cancer, liver cancer cell lines and general tissue of liver, finding bFGF were highly expressed in tissues of liver cancer and liver cancer cell lines. Affinity chromatography purifying bFGF was set up by mAb binging CNBr-pepharose 4B, and the purification was 95%. We found that the titer of anti-bFGF antibody was very high in serum of neuropathic amyotrophia.

应用细胞融合制备46株杂交瘤，对其中10株进行克隆化，获得bFGF特异单抗16株，2株针对融合蛋白；应用该单抗建立了0.1ng/孔灵敏度的ELISA，0.5ng/孔敏度的斑点ELISA；用Western-blotting鉴别了经改造不同工程菌蛋白表达，指导重组工程菌改造；用3T3细胞培养研究了单抗抑制bFGF生物学活性的剂量和特点；合作研究了20例肝癌、肝癌细胞株和正常肝组织，发现前者bFGF高表达；应用单抗偶联CNBr-sepharose 4B建立了小量免疫亲和层析纯化bFGF，纯度达到95%；发现神经性肌萎缩患者血清中含有高滴度的bFGF抗体，已有10多家单位引用单抗或进行合作。

Get high purity DCs by Cultured plastic-adherent monocytes isolated from healthy human peripheral blood with GM-CSF and IL-4 for 7 days. To observe the morphology of DCs by inverted phase contrast microscope ,electron microscope and laser confocal microscope. Analyse phenotype of DCs with flow cytometry. Investigate the endocytosis ability of DCs as a group by Horseradish peroxidase endocytosis assay. To appraise allogeneic mixed lymphocytes reaction of DCs by MTT reduction assay. Analyse the levels of IL-12 and TNF in liquids of cultured medium by ELISA and MTT reduction assay respectively. Soluble antigens of HCCs was obtained by 3 freeze-and-thaw cycles. Biological characteristics of HC soluble antigens pulsed DCs were monitored by flow cytometry. According to MTT reduction assay estimated the cell proliferation of self lymphocytes activated by HC antigens pulsed DCs. Get high purity BCG HSP 70 protein by SDS-PAGE electrophoresis and determined its biological activity with ELISA. Analyse phenotype of antigen pulsed DCs primed by BCG HSP70 with flow cytometry. By MTT reduction assay estimated the cell proliferation of self lymphocytes and the MLR of DC based vaccine. Analyse expression of HLA-DR molecule on surface of HCC lines. The IFN-γ mRNA in lymphocytes after actived by DC vaccine and the Fas-L expression on DC and DC vaccine primed lymphocytes were detected by in situ hybridization and flow cytometry respectively. Specific cytotoxity lysis of T lymphocytes and nonspecific inhibition of liquids in culture medium against HCC lines were also tested. Detect expression of hAFP on four HCC lines with Cell-ELISA. Induce apoptosis of HCCs with actinomycin-D. Interaction of DCs and apoptotic cells was observed under transmission electron microscope. Growth inhibition test of DC against HCC lines was also performed. Establish the nude mouse model bearing human HC xenografts and indentify the characteristic of tumour by histochemistry and immunohistochemistry techniques. Prevent and treat transplanted human HC on nude mouse with Freezing and anabiotic HC specific lymphocytes.

用GM-CSF和IL-4从健康人外周血诱导DC；分别用倒置相差显微镜、电子显微镜及激光共聚焦显微镜观察DC形态；流式细胞术检测DC表型；HRP吞噬实验测定DC的群体内吞能力；MTT法检测同种异体混合淋巴细胞反应；ELISA法和MTT法分别测定DC培养上清液中IL-12和TNF水平；冻融法制备肝癌细胞可溶性抗原；流式细胞术检测负载肝癌可溶性抗原后DC的生物学特性；MTT法检测DC负载肝癌抗原后对自身淋巴细胞增殖的影响；SDS-PAGE制备电泳纯化BCG HSP70并鉴定纯度，ELISA测定活性；流式细胞术检测负载抗原DC经BCGHSP 70活化后的表型；MTT法检测肝癌DC疫苗对自身淋巴细胞增殖的影响和混合淋巴细胞反应；流式细胞术检测肝癌细胞表面HLA-DR表达；MTT法检测肝癌DC疫苗对自身淋巴细胞的活化；原位杂交法检测肝癌DC疫苗活化后的淋巴细胞IFN-γmRNA表达；流式细胞术检测DC和肝癌DC疫苗活化后淋巴细胞表面Fas-L；MTT法分别检测肝癌DC疫苗活化的淋巴细胞和其培养上清对肝癌细胞的特异性杀伤和非特异性抑制作用；Cell-ELISA检测人肝癌细胞hAFP表达；MTT法检测负载AFP表位肽和凋亡肝癌细胞DC对自身淋巴细胞增殖的影响；ELISA法和MTT法分别测定活化后淋巴细胞培养上清中TNF和IL-12水平；肝癌细胞凋亡的诱导和检测；DC吞噬凋亡肝癌细胞后的电子显微镜观察；DC对肝癌细胞的生长抑制试验；人肝癌裸鼠皮下移植瘤动物模型的建立及其组织学和免疫组织化学鉴定；DC及肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤；冻存和复苏后的肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤。

The method takes microcrystalline cellulose as affinity column chromatographic medium of Beta-glucosaccharase; under low temperature and anacid condition, the sample of Beta-glucosaccharase is loaded and specifically adsorbed to the microcrystalline column, and then the concentration of NaCl is increased to elute the enzyme, so as to achieve high-efficient and economic separation and purification.

本方法以微晶纤维素作为β-葡萄糖苷酶的亲和柱层析介质，在低温和微酸性条件下，将β-葡萄糖苷酶上样并特异性吸附于微晶纤维素柱，然后再通过增加NaCl浓度的方法将该酶洗脱，从而达到其高效而经济分离纯化的目的。

Crude extract of GCA went through deproteinization, depigmentation, ethanol reclamation, salting-out, extraction and anhydration.

采用除蛋白、除色素、回收乙醇、盐析、萃取等生物化学方法，从羊胆汁中分离提取GCA，采用了吸附色谱法和薄层层析色谱法分离纯化和鉴定了GCA。

With the SDS-PAGE preparation electrophoresis, high quality insecticidal protein was purified from a unique Bacillus thuringiensis serovar japonensis strain HBF-1, which is highly toxic to larvae of Anomala exoleta and A.

以苏云金芽孢杆菌HBF-1菌株中分离纯化的高特异性杀虫蛋白作为抗原，免疫新西兰白兔制备出多克隆抗体，其效价可达到16000。

We isolate the LCA binding glycosylated proteins from anosia human liver cell extraction and liver cancer cell extraction by LCA coupled epoxy magnetic particles, and the glycosylated fractions were separated by 2D gels which were analyzed by 2D ImageMaster to established disparity expression spectra between Chang Liver and SMMC-7721 firstly.

利用该方法对健康人肝脏细胞Chang Liver总蛋白、肝癌细胞SMMC-7721总蛋白及健康人血清进行了糖蛋白的分离纯化，结合2D电泳技术及其图像分析，建立了健康人肝脏细胞ChangLiver和肝癌细胞SMMS-7721 LCA结合糖蛋白的差异表达谱，分别得到蛋白质点148±5和209±7个，匹配点数为127，匹配率为85.36%。

The thesis is concerned with the fermentation of yeast Candida arborea AS2.566, separation and purification of L-tryptophan and synthesis of 15N labeled anthranilic acid.

本文以发酵法生产L-色氨酸为背景，重点研究了由假丝酵母AS2.566获得L-色氨酸的发酵条件、色氨酸的分离纯化工艺以及邻氨基苯甲酸的合成过程。

SR-B, a colorless crystal antimicrobic compound, was extracted and purified from broth of marine actinomycete N331 isolated and screened from sea mud. Its molecular structure was characterized by MS and various NMR spectra.

从分离筛选的海洋放线菌N331的发酵液中分离、纯化得到一种无色结晶抗菌化合物SR-B，通过对其质谱、多种核磁共振谱的结构信息进行分析，鉴定其分子结构。

Chinese olive as raw material, study its antioxygenic activity.

以青橄榄为原料，提取纯化青橄榄黄酮并研究其抗氧化活性。

As she looked at Warrington's manly face, and dark, melancholy eyes, she had settled in her mind that he must have been the victim of an unhappy attachment.

每逢看到沃林顿那刚毅的脸，那乌黑、忧郁的眼睛，她便会相信，他一定作过不幸的爱情的受害者。

Maybe they'll disappear into a pothole.

也许他们将在壶穴里消失