Hybridoma and hybridomics最新文献

The clinical significance of molecular markers to bladder cancer. 分子标志物对膀胱癌的临床意义。

Hybridoma and hybridomics Pub Date : 2004-12-01 DOI: 10.1089/hyb.2004.23.335

Konstantinos N Syrigos, Eleni Karapanagiotou, Kevin J Harrington

{"title":"The clinical significance of molecular markers to bladder cancer.","authors":"Konstantinos N Syrigos, Eleni Karapanagiotou, Kevin J Harrington","doi":"10.1089/hyb.2004.23.335","DOIUrl":"https://doi.org/10.1089/hyb.2004.23.335","url":null,"abstract":"Stage and grade of transitional cell carcinoma are currently the most useful tools for taking therapeutic decisions and evaluating the prognosis of bladder cancer patients. However, as there are remarkable differences in biological behavior and \"biological potential\" of tumors classified in the same stage, it is very difficult to predict which superficial tumors will recur and which tumors will give distant metastases. During the last two decades, the better understanding of the molecular mechanisms involved in carcinogenesis and tumor progression has provided a large number of molecular markers of bladder cancer, with a potential diagnostic and prognostic value. This article reviews comprehensively the molecular role and evaluates the clinical significance and the perspectives of these molecular markers. We concluded that, although at the moment there is not a single marker able to predict with accuracy the biological potential of bladder cancer, the most promising markers, at this point, are deletions of chromosome 9, and the tumor suppressor gene p53. Clinical studies are in progress for the assessment of other biological molecules with prognostic potential, such as the E-cadherin, the protein p120, and the telomerase.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"23 6","pages":"335-42"},"PeriodicalIF":0.0,"publicationDate":"2004-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/hyb.2004.23.335","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24935349","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 22

Inhibition of Porphyromonas gingivalis hemagglutinating activity by synthetic peptides derived from phage display selection using MAb against the recombinant outer membrane protein. 噬菌体展示选择合成肽对重组外膜蛋白的抑制作用。

Hybridoma and hybridomics Pub Date : 2004-12-01 DOI: 10.1089/hyb.2004.23.352

Wei-Jen Chang, Michiko Kishikawa-Kiyama, Yasuko Shibata, Sheng-Yang Lee, Yoshimitsu Abiko

{"title":"Inhibition of Porphyromonas gingivalis hemagglutinating activity by synthetic peptides derived from phage display selection using MAb against the recombinant outer membrane protein.","authors":"Wei-Jen Chang, Michiko Kishikawa-Kiyama, Yasuko Shibata, Sheng-Yang Lee, Yoshimitsu Abiko","doi":"10.1089/hyb.2004.23.352","DOIUrl":"https://doi.org/10.1089/hyb.2004.23.352","url":null,"abstract":"Porphyromonas gingivalis has been implicated as an pathogen in the development of periodontitis, and hemagglutinins have been identified as an important adhesion onto the gingival tissue cells, and to attach and lyse erythrocytes to uptake Fe ion as an essential nutriant. The 40-kDa outer membrane protein (OMP) has been moleculary cloned from P. gingivalis 381. Since the antibody against recombinant (r) 40-kDa OMP inhibited the hemagglutinating activity, and the polymeric form of r40-kDa OMP itself expressed hemagglutinating activity, the 40-kDa OMP is thought to be one of the hemagglutinins. Moreover, we established MAbs against r40-kDa OMP which were capable of inhibiting hemagglutinating activity of P. gingivalis vesicles. In the present study, a phage-displayed epitope mapping system was used to identify the functional domain expressing hemagglutinating activity by biopanning using the neutralizing mAb, Pg-ompA1. The minimal epitope requirements of the MAb and the predicted amino acid sequences were identified in the region of (96)IALDQTLGIP(105) in 40-kDa OMP. Synthetic peptide, (87)WPRVGQLFIALDQTLGIPTFSVCRME(116), mapped the relevant molecule within a short stretch and is corresponding to residues of 40-kDa OMP. Chemically synthesized peptide was used to determine its inhibitory activity against hemagglutinating activity. The synthetic peptide significantly abolished hemagglutinating activity in a dose-dependent manner. These findings suggest that the synthetic peptide is an effective antagonist of erythrocyte binding, and this peptide may be a potent inhibitor of hemagglutination of P. gingivalis cells. The use of synthetic peptide neutralizing hemagglutinating activity of P. gingivalis represents a possible new therapeutic approach to P. gingivalis infected periodontitis.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"23 6","pages":"352-6"},"PeriodicalIF":0.0,"publicationDate":"2004-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/hyb.2004.23.352","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24935351","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10

Diagnostic application of immunoperoxidase monolayer assay using monoclonal antibodies produced against equine arteritis virus 14-kDa nucleocapsid protein. 马动脉炎病毒14-kDa核衣壳蛋白单克隆抗体免疫过氧化物酶单层检测在诊断中的应用

Hybridoma and hybridomics Pub Date : 2004-12-01 DOI: 10.1089/hyb.2004.23.368

Akos Hornyák, Béla Dénes, Levente Szeredi, László Dencsö, Miklós Rusvai

引用次数: 4

Serine and threonine phospho-specific antibodies to p120-catenin. p120-catenin丝氨酸和苏氨酸磷酸化特异性抗体。

Hybridoma and hybridomics Pub Date : 2004-12-01 DOI: 10.1089/hyb.2004.23.343

Xiaobo Xia, James Brooks, Roberto Campos-González, Albert B Reynolds

引用次数: 18

Sensitive detection of human IgG in ELISA using a monoclonal anti-IgG-peroxidase conjugate. 单克隆抗IgG-过氧化物酶偶联物ELISA灵敏检测人IgG。

Hybridoma and hybridomics Pub Date : 2004-12-01 DOI: 10.1089/hyb.2004.23.362

Marie-Claire Chevrier, Isabel Châteauneuf, Matthieu Guérin, Réal Lemieux

{"title":"Sensitive detection of human IgG in ELISA using a monoclonal anti-IgG-peroxidase conjugate.","authors":"Marie-Claire Chevrier, Isabel Châteauneuf, Matthieu Guérin, Réal Lemieux","doi":"10.1089/hyb.2004.23.362","DOIUrl":"https://doi.org/10.1089/hyb.2004.23.362","url":null,"abstract":"Enzyme-antibody (Ab) conjugates specific for IgG are widely used in indirect immunological assays and have been until recently routinely prepared with polyclonal IgG-specific animal Abs. The use of monoclonal Abs (MAbs) could permit a better standardization of the IgG-specific conjugate reagents but is expected to result in lower reactivity due to the recognition of a single epitope by the MAbs. In this work, we have characterized a monoclonal anti-human IgG-peroxidase (HRP) reagent and compared its reactivity with commercial reagents. The murine C5-1 anti-human IgG MAb was selected for conjugation because of its high affinity (K(a) = 1.9 x 10(10)M), pan-IgG reactivity and absence of cross-reactivity with various structures including animal IgGs. The specific activity and binding kinetics of the C5-1:HRP conjugate were similar to the ones of two polyclonal anti-IgG:HRP conjugates when tested with immobilized human IgG. The C5-1:HRP conjugate could detect low amounts of human IgG much more effectively than two commercial monoclonal conjugates although it was slightly less effective than a polyclonal conjugate. However, the C5-1 conjugate yielded reduced background reactivity compared to the polyclonal conjugate, resulting in similar signal-to-noise ratios. These results indicate that the C5-1:HRP conjugate could be a suitable substitute for anti-human IgG conjugates prepared from animal antisera.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"23 6","pages":"362-7"},"PeriodicalIF":0.0,"publicationDate":"2004-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/hyb.2004.23.362","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24935353","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 11

Isolation and characterization of the human monoclonal antibodies C10 in single-chain fragment variable (scFv) format to glucose oxidase from Aspergillus niger. 黑曲霉葡萄糖氧化酶单克隆抗体C10单链片段变量(scFv)的分离与鉴定

Hybridoma and hybridomics Pub Date : 2004-12-01 DOI: 10.1089/hyb.2004.23.380

Alessandro Ascione, Michela Flego, Silvia Zamboni, Emanuela De Cinti, Maria Luisa Dupuis, Maurizio Cianfriglia

{"title":"Isolation and characterization of the human monoclonal antibodies C10 in single-chain fragment variable (scFv) format to glucose oxidase from Aspergillus niger.","authors":"Alessandro Ascione, Michela Flego, Silvia Zamboni, Emanuela De Cinti, Maria Luisa Dupuis, Maurizio Cianfriglia","doi":"10.1089/hyb.2004.23.380","DOIUrl":"https://doi.org/10.1089/hyb.2004.23.380","url":null,"abstract":"Despite biotechnological and clinical applications very few monoclonal antibodies (MAbs) directed to the enzyme glucose oxidase, have been produced so far because of the heavy side effects of the immunization schedule for conventional MAb preparation. In contrast, the phage display method allows for the selection of monoclonal human antibody fragments against any antigens, including toxic proteins. Furthermore, cDNA sequences encoding selected antibody fragments are readily identified, facilitating various molecular targeting approaches. In order to obtain such human fragments recognizing glucose oxidase, we used the large synthetic ETH-2 library based on the principle of protein design. Phage displaying glucose oxidase reactive scFvs were obtained after three rounds of selection on glucose oxidase-coated immunotubes and subsequent amplification in TG1 E. coli cells. Eventually, one high reactive scFv clone was selected and further examined. The anti-glucose oxidase scFv C10 was found suitable for Western blot; Biacore analysis showed that the binding affinity of the glucose oxidase-reactive scFv is almost equal that of MAbs prepared with conventional hybridoma technology. Finally, the cDNA sequence of this human scFv may be exploited to generate bispecific antibodies to target in the tumor environment-specific toxic enzymatic reaction.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"23 6","pages":"380-4"},"PeriodicalIF":0.0,"publicationDate":"2004-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/hyb.2004.23.380","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24935356","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 15

Generation of monoclonal antibodies against mouse neurogenin 3: a new immunocytochemical tool to study the pancreatic endocrine progenitor cell. 小鼠神经原素3单克隆抗体的制备:研究胰腺内分泌祖细胞的一种新的免疫细胞化学工具。

Hybridoma and hybridomics Pub Date : 2004-12-01 DOI: 10.1089/hyb.2004.23.385

Stefan Zahn, Jakob Hecksher-Sørensen, Inger Lund Pedersen, Palle Serup, Ole Madsen

{"title":"Generation of monoclonal antibodies against mouse neurogenin 3: a new immunocytochemical tool to study the pancreatic endocrine progenitor cell.","authors":"Stefan Zahn, Jakob Hecksher-Sørensen, Inger Lund Pedersen, Palle Serup, Ole Madsen","doi":"10.1089/hyb.2004.23.385","DOIUrl":"https://doi.org/10.1089/hyb.2004.23.385","url":null,"abstract":"The induction of pancreatic endocrine differentiation from undifferentiated precursor cells is a multi-step process regulated by the expression of several transcription factors. At E9.5 expression of homeodomain protein Pdx-1 defines the early pancreatic epithelium. Later, expression of the basic helix-loop-helix factor, Neurogenin 3 (Ngn3) marks the initiation of the endocrine lineage from the pancreatic precursor cells. A full understanding of these processes is essential in order to control development of insulin secreting beta-cells from embryonic stem cells in vitro. Since the expression of ngn3 is a key step, the aim of this work was to develop monoclonal antibodies for immunohistochemical (IHC) detection of Ngn3. All mice (RBF-strain) immunized with recombinant GST-mNGN3 fusion protein responded with a high antibody titer. The sera were further screened by IHC on paraffin sections of fetal (E13) mouse pancreas, and two hybridomas subsequently derived from one selected mouse produced antibodies with prominent and specific Ngn3 staining properties. These new antibodies provide novel useful tools for co-labeling studies using the large panel of existing rabbit polyclonal antibodies available against important transcription factors to further characterize the development of pancreatic endocrine progenitor cells.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"23 6","pages":"385-8"},"PeriodicalIF":0.0,"publicationDate":"2004-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/hyb.2004.23.385","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24935299","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 23

Preparation and characterization of monoclonal antibody against abalone allergen tropomyosin. 抗鲍鱼过敏原原肌球蛋白单克隆抗体的制备与鉴定。

Hybridoma and hybridomics Pub Date : 2004-12-01 DOI: 10.1089/hyb.2004.23.357

Ying Lu, Toshiaki Oshima, Hideki Ushio, Kazuo Shiomi

{"title":"Preparation and characterization of monoclonal antibody against abalone allergen tropomyosin.","authors":"Ying Lu, Toshiaki Oshima, Hideki Ushio, Kazuo Shiomi","doi":"10.1089/hyb.2004.23.357","DOIUrl":"https://doi.org/10.1089/hyb.2004.23.357","url":null,"abstract":"Muscle protein tropomyosin is a major and common allergen of mollusks. In order to develop immunoassays based on monoclonal antibody (MAb) for allergen characterization, a MAb against Japanese abalone (Haliotis discus) was prepared and characterized in the present study. In comparison with the IgE reactivities of sera from crustacean allergic individuals, the selected MAb AE9F9 showed specific reaction to the abalone allergenic tropomyosin. The MAb AE9F9 reacted to the crustaceans including lobster, crab, and shrimp, but not to the mollusks other than abalone. It was surprising that the MAb AE9F9 was also reactive to the vertebrate chicken tropomyosin and smooth muscle tropomyosin of chicken gizzard. Comparison of amino acid sequences of tropomyosins among abalone, other mollusks, crustaceans, and chicken showed that two regions (90-105 and 147-165) have a high identity among abalone, crustaceans, and chicken, but are polymorphic among mollusks. In the two regions, substitutions at residues 99-Leu, 149-Lys, and 160-Arg of abalone tropomyosin are observed only in mollusks, suggesting that these residues might be important for determining the abalone tropomyosin structure recognized by the AE9F9 antibody. Because of the distinct binding site, the MAb AE9F9 might be useful for abalone allergen detection and epitope mapping of allergen tropomyosin.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"23 6","pages":"357-61"},"PeriodicalIF":0.0,"publicationDate":"2004-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/hyb.2004.23.357","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24935352","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 13

Development of a quantitative cell-based intracellular ELISA for the screening of B cell hybridoma supernatants: a novel rapid assay to detect positive clones. B细胞杂交瘤上清液筛选的细胞内定量ELISA方法的建立:一种检测阳性克隆的新型快速测定方法。

Hybridoma and hybridomics Pub Date : 2004-12-01 DOI: 10.1089/hyb.2004.23.373

J Gourapura Renukaradhya, Venkataraman Sriram, Katarina Polakova, Gustav Russ, Randy R Brutkiewicz

引用次数: 6

Expression system for enhanced green fluorescence protein conjugated recombinant antibody fragment. 增强型绿色荧光蛋白偶联重组抗体片段的表达系统。

Hybridoma and hybridomics Pub Date : 2004-10-01 DOI: 10.1089/hyb.2004.23.279

Kye Sook Yi, Junho Chung, Kwang-Hyun Park, Kisu Kim, Shin-Young Im, Cha-Yong Choi, Mie-Jae Im, Uh-Hyun Kim

{"title":"Expression system for enhanced green fluorescence protein conjugated recombinant antibody fragment.","authors":"Kye Sook Yi, Junho Chung, Kwang-Hyun Park, Kisu Kim, Shin-Young Im, Cha-Yong Choi, Mie-Jae Im, Uh-Hyun Kim","doi":"10.1089/hyb.2004.23.279","DOIUrl":"https://doi.org/10.1089/hyb.2004.23.279","url":null,"abstract":"Recent development of recombinant antibody technology has enabled fusion of recombinant antibody fragment with fluorescent proteins for various applications such as flow cytometry, fluorescence immunoassay, and fluorescent microscopy. In this study, we generated various forms of green fluorescence protein (EGFP)-fused anti-c-Met antibody fragment. Among these fusion proteins, EGFP fusion to the light chain showed high expression in a soluble form of protein in E. coli, and high binding activity to c-Met. A feasibility of the constructs was further examined by replacing the Fab gene by a Fab library of catalytic subunit of protein kinase A (PKA) to construct the Fab library in EGFP fused form. We also constructed the conventional Fab library. After a series of biopanning, we found that the binding capability of EGFP-anti-PKA Fab was comparable with anti-PKA Fab. Sequence analysis of the selected clones showed > or =99% identity in amino acid sequence and shared the same CDR sequence. These results demonstrate that EGFP fusion to the light chain using our vector system does not influence the selection of reactive Fab and that this vector system is useful for EGFP fusion to Fab to develop a one-step detection system.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"23 5","pages":"279-86"},"PeriodicalIF":0.0,"publicationDate":"2004-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/hyb.2004.23.279","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"24924822","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 10