Hybridoma and hybridomics最新文献_第10页

Immunological characterization of exocyst complex subunits in cell differentiation. 胞囊复合体亚基在细胞分化中的免疫学特性。

Hybridoma and hybridomics Pub Date : 2003-06-01 DOI: 10.1089/153685903322286575

Sheng Wang, Shu C Hsu

引用次数: 10

Minimal structural elements of an inhibitory anti-ATF1/CREB single-chain antibody fragment (scFv41.4). 抑制抗atf1 /CREB单链抗体片段(scFv41.4)的最小结构元素。

Hybridoma and hybridomics Pub Date : 2003-04-01 DOI: 10.1089/153685903321947987

R J Olsen, J Mazlo, S A Koepsell, T W McKeithan, S H Hinrichs

{"title":"Minimal structural elements of an inhibitory anti-ATF1/CREB single-chain antibody fragment (scFv41.4).","authors":"R J Olsen, J Mazlo, S A Koepsell, T W McKeithan, S H Hinrichs","doi":"10.1089/153685903321947987","DOIUrl":"https://doi.org/10.1089/153685903321947987","url":null,"abstract":"Antibody variable domains represent potential structural models for the rational design of therapeutic molecules that bind cellular proteins with high affinity and specificity. The Activating Transcription Factor 1 (ATF1)/Cyclic AMP Response Element Binding Protein (CREB) family of transcription factors are particularly relevant targets due to their strong association with melanoma and clear cell sarcoma. Biochemical and structural investigations were performed to optimize a single-chain antibody fragment (scFv), scFv41.4, that disrupts the binding of ATF1/CREB to cyclic-AMP response elements (CRE) in vitro and inhibits transcriptional activation in cells. Molecular modeling and ligand docking simulations suggested that scFv41.4 could function as a disulfide-deficient single domain scFv. Functional studies verified that deletion of the light chain did not result in reduced inhibitory activity. The isolated heavy chain was predicted to assume a relaxed structural conformation that maintained a functional antigen binding pocket. The minimal structural elements necessary for intracellular function were further analyzed by selective deletion of CDR1 and CDR2. V(H)-CDR1 and V(H)-CDR3 were shown to play a key role in antigen binding activity, but V(H)-CDR2 was dispensable. Thus, scFv41.4 represents a unique molecule with potential for use in the design of peptidomimetic derivatives having therapeutic application to human cancer.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"22 2","pages":"65-77"},"PeriodicalIF":0.0,"publicationDate":"2003-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/153685903321947987","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22459159","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: 4

A panel of monoclonal antibodies to cytoplasmic GW bodies and the mRNA binding protein GW182. 细胞质GW小体和mRNA结合蛋白GW182的单克隆抗体。

Hybridoma and hybridomics Pub Date : 2003-04-01 DOI: 10.1089/153685903321947996

Theophany Eystathioy, Edward K L Chan, Michael Mahler, Leeanne M Luft, Mark L Fritzler, Marvin J Fritzler

{"title":"A panel of monoclonal antibodies to cytoplasmic GW bodies and the mRNA binding protein GW182.","authors":"Theophany Eystathioy, Edward K L Chan, Michael Mahler, Leeanne M Luft, Mark L Fritzler, Marvin J Fritzler","doi":"10.1089/153685903321947996","DOIUrl":"https://doi.org/10.1089/153685903321947996","url":null,"abstract":"GW182 is a mRNA binding protein characterized by 60 repeats of glycine (G):tryptophan (W) motifs and is localized in cytoplasmic structures referred to as GW bodies (GWBs). Current evidence suggests that this unique protein plays a role in mRNA processing. To enable a more detailed study of GW182 and GWBs in cells and tissues, including their role in mRNA processing, we developed four monoclonal antibodies (MAbs) that bind the human recombinant GW182 protein. These MAbs can be used for Western blot analysis and indirect immunofluorescence (IIF) on cultured cells and tissues. Of special interest, one of the MAbs, 2D6, can be used to identify GW182 and GWBs in formalin-fixed and paraffin-embedded tissues after using an antigen retrieval method (ARM). All the MAbs described in this study immunoprecipitate the GW182 protein. Epitope mapping using overlapping 15-mer peptides representing the full-length GW182 showed that the major antibody-binding domains of these MAbs are distinct. These MAbs are valuable tools for cell biologists and pathologists to study the location and function of the novel GW182 protein in tissue culture cells, as well as cryopreserved or archived tissues.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"22 2","pages":"79-86"},"PeriodicalIF":0.0,"publicationDate":"2003-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/153685903321947996","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22458604","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: 34

Preparation of humanized ovarian carcinoma anti-idiotypic minibody. 人源化卵巢癌抗独特型小体的制备。

Hybridoma and hybridomics Pub Date : 2003-04-01 DOI: 10.1089/153685903321948030

Xiaohong Chang, Heng Cui, Jie Feng, Yi Li, Bei Liu, Shanjin Cao, Yexia Cheng, Henian Qian

{"title":"Preparation of humanized ovarian carcinoma anti-idiotypic minibody.","authors":"Xiaohong Chang, Heng Cui, Jie Feng, Yi Li, Bei Liu, Shanjin Cao, Yexia Cheng, Henian Qian","doi":"10.1089/153685903321948030","DOIUrl":"https://doi.org/10.1089/153685903321948030","url":null,"abstract":"Murine anti-idiotypic monoclonal antibody (MAb) 6B11 mimicking the tumor-associated antigen OC166-9 is used as a vaccine for the induction of an anti-tumoral immunity in experiments of in vitro and in vivo animal model with ovarian carcinoma. In this article, we have humanized 6B11 anti-idiotypic minibody using overlap polymerase chain reaction (PCR) and DNA recombinant technique, prokaryotic expression vector was produced by genetic fusion of 6B11V(L)-V(H) to human IgG1 hinge and CH3 region. Transformed E. coli BL21(DE3) were propagated and induced by isopropyl-beta D-thiogalactopyranoside (IPTG). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that a protein band with molecular weight of 50kD appeared as the expected size after transformation. Molecular weight of 100 kDa may be examined by electrophoresis in nondenaturing systems. The fusion protein was analyzed with enzyme-linked immunosorbant assay (ELISA), inhibition ELISA tests and Western blot, respectively. The humanized anti-idiotype minibody showed capacity of bivalent binding to ovarian cancer MAb COC166-9 and goat anti-human immunoglobulin IgG1. It is useful reagents for clinical use.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"22 2","pages":"109-15"},"PeriodicalIF":0.0,"publicationDate":"2003-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/153685903321948030","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22458608","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: 9

1B11, 1C7, 2C5, 2C9, 3G12, 4H9, 5B8, 5C11, 5D3, 5E5, 6F11, and 6H10 Anti-Mycoplasma bovis 抗牛支原体1B11、1C7、2C5、2C9、3G12、4H9、5B8、5C11、5D3、5E5、6F11、6H10

Hybridoma and hybridomics Pub Date : 2003-04-01 DOI: 10.1089/153685903321948085

B. Dénes

引用次数: 0

Development of functional human monoclonal single-chain variable fragment antibody against HIV-1 from human cervical B cells. 人宫颈B细胞抗HIV-1单克隆单链可变片段抗体的制备。

Hybridoma and hybridomics Pub Date : 2003-04-01 DOI: 10.1089/153685903321948021

Jody D Berry, John Rutherford, Gregg J Silverman, Rupert Kaul, Marikka Elia, Sarah Gobuty, Roberta Fuller, Francis A Plummer, Carlos F Barbas

{"title":"Development of functional human monoclonal single-chain variable fragment antibody against HIV-1 from human cervical B cells.","authors":"Jody D Berry, John Rutherford, Gregg J Silverman, Rupert Kaul, Marikka Elia, Sarah Gobuty, Roberta Fuller, Francis A Plummer, Carlos F Barbas","doi":"10.1089/153685903321948021","DOIUrl":"https://doi.org/10.1089/153685903321948021","url":null,"abstract":"A panel of novel recombinant single-chain variable fragment (scFv) antibody against human immunodeficiency virus type-1 (HIV-1) was isolated and characterized. We generated human scFvs using RNA harvested from cervical B lymphocytes of Kenyan prostitutes who are highly exposed to HIV-1, but remain persistently seronegative. The variable regions of the heavy (VH) and light (VL) chain antibody genes were selected as hybrids using guided-selection with the VL and VH, respectively, of a derivative of IgGb(12) using the phagemid vector pComb3X. IgGb(12) is a previously well-characterized HIV-1 neutralizing human monoclonal antibody (MAb). One of the hybrid scFv, IgA6/4L, neutralizes HIV-1 infectivity in in vitro cell culture assay. The cervical VH and VL chain antibody genes were connected by a DNA linker and subcloned in pComb3X. The cervical scFv clones were functional in recognizing HIV-1 gp120 by enzyme-linked immunosorbant assay (ELISA) and on cells in flow cytometry. Whole IgGb(12) does not inhibit binding of clones IgA6/5k nor IgA6/30lambda to gp120, which suggests that they bind different epitopes. Nucleotide sequence analysis of the cervical scFv show the clones are unique and reveal interesting characteristics of human cervical V gene pools. This work demonstrates, for the first time, cloning of a functional scFv MAb to a sexually transmitted disease pathogen from local cervical B-cell pools in exposed humans.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"22 2","pages":"97-108"},"PeriodicalIF":0.0,"publicationDate":"2003-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/153685903321948021","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22458607","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

Preparation and characterization of a set of monoclonal antibodies to TRAIL and TRAIL receptors DR4, DR5, DcR1, and DcR2. TRAIL和TRAIL受体DR4、DR5、DcR1和DcR2单克隆抗体的制备和鉴定

Hybridoma and hybridomics Pub Date : 2003-04-01 DOI: 10.1089/153685903321948058

Xue-Song Liu, Yong Zhu, Wei-Ning Han, Ying-Na Li, Li-Hua Chen, Wei Jia, Chao-Jun Song, Fei Liu, Kun Yang, Qi Li, Bo-Quan Jin

{"title":"Preparation and characterization of a set of monoclonal antibodies to TRAIL and TRAIL receptors DR4, DR5, DcR1, and DcR2.","authors":"Xue-Song Liu, Yong Zhu, Wei-Ning Han, Ying-Na Li, Li-Hua Chen, Wei Jia, Chao-Jun Song, Fei Liu, Kun Yang, Qi Li, Bo-Quan Jin","doi":"10.1089/153685903321948058","DOIUrl":"https://doi.org/10.1089/153685903321948058","url":null,"abstract":"The tumor necrosis factor-related apoptosis-inducing ligand (TRAIL/Apo 2L) is a novel cytotoxic ligand belonging to TNF superfamily. Among TRAIL receptors, death receptor 4 (DR4) and DR5 containing death domain (DD) in their cytoplasmic region mediate apoptosis-signaling upon TRAIL binding, while decoy receptor 1 (DcR1) and DcR2 with a truncated or non-functional DD play \"decoy\" role. The interaction of TRAIL and TRAIL receptors plays important roles both in immunoregulation and immune pathogenesis of some diseases. In this study, we raised hybridomas secreting monoclonal antibodies against TRAIL (FMU1.1, 1.2, 1.3), DR4 (FMU1.4), DR5 (FMU1.5, 1.6), DcR1 (FMU1.7) and DcR2 (FMU1.8, 1.9). These MAbs could be used for fluorescent staining and flow cytometry (FCM) analysis as well as immunohistochemistry (IHC). Moreover, FMU1.1, 1.3, 1.4 and 1.5 could be used as coating antibodies paring corresponding polyclonal antibodies to develop sandwich ELISAs to quantitate the soluble TRAIL (sTRAIL), sDR4 or sDR5 in serum samples respectively. In addition, cross-linking of DR4/DR5 by FMU1.4 or FMU1.5 MAbs could induce apoptosis of some DR4/DR5-expressing tumor cells. Thus, this set of monoclonal antibodies against TRAIL or TRAIL receptors may be useful in expression phenotypic and functional study of TRAIL and TRAIL receptors.","PeriodicalId":83733,"journal":{"name":"Hybridoma and hybridomics","volume":"22 2","pages":"121-5"},"PeriodicalIF":0.0,"publicationDate":"2003-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1089/153685903321948058","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"22458610","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: 5

Generation and characterization of species-specific anti-Bcl-X(L) monoclonal antibodies. 物种特异性抗bcl - x (L)单克隆抗体的制备和鉴定。

Hybridoma and hybridomics Pub Date : 2003-04-01 DOI: 10.1089/153685903321948012

Yi-Te Hsu, Qi Hou, Eugene Cymbalyuk, Richard Youle

引用次数: 3

Generation of a monoclonal antibody against the mouse Sf3b1 (SAP155) gene product for U2 snRNP component of spliceosome. 制备了一种针对小鼠Sf3b1 (SAP155)基因产物的U2 snRNP剪接体单克隆抗体。

Hybridoma and hybridomics Pub Date : 2003-04-01 DOI: 10.1089/153685903321948049

Atsuya Horie, Kyoichi Isono, Haruhiko Koseki

引用次数: 1

Production of monoclonal antibodies recognizing human hair follicle keratinocytes. 识别人毛囊角质形成细胞单克隆抗体的制备。

Hybridoma and hybridomics Pub Date : 2003-04-01 DOI: 10.1089/153685903321948067

Shin Hatakeyama, Ying-Zhi Ma, Naoko Miura, Shoko Abe, Takashi Kameda, Kenji Sakamoto, Toshihiro Sugiyama

引用次数: 1