Berry JD et al. (SEP 2004)
Journal of Virological Methods 120 1 87--96
Development and characterisation of neutralising monoclonal antibody to the SARS-coronavirus
There is a global need to elucidate protective antigens expressed by the SARS-coronavirus (SARS-CoV). Monoclonal antibody reagents that recognise specific antigens on SARS-CoV are needed urgently. In this report,the development and immunochemical characterisation of a panel of murine monoclonal antibodies (mAbs) against the SARS-CoV is presented,based upon their specificity,binding requirements,and biological activity. Initial screening by ELISA,using highly purified virus as the coating antigen,resulted in the selection of 103 mAbs to the SARS virus. Subsequent screening steps reduced this panel to seventeen IgG mAbs. A single mAb,F26G15,is specific for the nucleoprotein as seen in Western immunoblot while five other mAbs react with the Spike protein. Two of these Spike-specific mAbs demonstrate the ability to neutralise SARS-CoV in vitro while another four Western immunoblot-negative mAbs also neutralise the virus. The utility of these mAbs for diagnostic development is demonstrated. Antibody from convalescent SARS patients,but not normal human serum,is also shown to specifically compete off binding of mAbs to whole SARS-CoV. These studies highlight the importance of using standardised assays and reagents. These mAbs will be useful for the development of diagnostic tests,studies of SARS-CoV pathogenesis and vaccine development. ?? 2004 Elsevier B.V. All rights reserved.
View Publication
产品号#：
03800
03801
03802
03803
03804
03805
03806
产品名：
ClonaCell™-HY杂交瘤试剂盒
ClonaCell™-HY培养基A
ClonaCell™-HY 培养基 B
ClonaCell™-HY 培养基 C
ClonaCell™-HY 培养基 D
ClonaCell™-HY 培养基 E
ClonaCell™-HY PEG
Kuroki MM et al. ( 2005)
Anticancer Research 25 6A 3733--9
Preparation of human IgG and IgM monoclonal antibodies for MK-1/Ep-CAM by using human immunoglobulin gene-transferred mouse and gene cloning of their variable regions.
For antibody-based therapy of cancer,monoclonal antibodies (mAbs) of human origin are superior to mouse,mouse/human chimeric or humanized mAbs,because of their minimum immunogenicity to humans and their efficient collaboration with human effector cells. In the present study,human mAbs were prepared against a pancarcinoma antigen,MK-1 (Ep-CAM),using a genetically-engineered mouse (KM mouse) that contains the human immunoglobulin genes. Spleen cells from KM mice,immunized with recombinant MK-1,were fused with P3-U1 mouse myeloma cells. Of 44 anti-MK-1 clones analyzed,two were of IgG4 and the others of IgM clones. Although the two IgG4 clones were suggested to recognize the same antigenic determinant or two closely located determinants,their VK regions were encoded by different light-chain genes while their VH sequences were identical. The two IgG4 and one of the IgM clones tested revealed antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity,respectively,against MK-1-expressing cells in vitro,suggesting that these fully human mAbs produced against MK-1 and their V-region genes,which are applicable for the preparation of engineered antibody fragments that may be useful for antibody-based therapy of cancer.
View Publication
产品号#：
03800
03801
03802
03803
03804
03805
03806
产品名：
ClonaCell™-HY杂交瘤试剂盒
ClonaCell™-HY培养基A
ClonaCell™-HY 培养基 B
ClonaCell™-HY 培养基 C
ClonaCell™-HY 培养基 D
ClonaCell™-HY 培养基 E
ClonaCell™-HY PEG
Wilson JR et al. (NOV 2016)
Antiviral Research 135 48--55
An influenza A virus (H7N9) anti-neuraminidase monoclonal antibody with prophylactic and therapeutic activity in vivo
Zoonotic A(H7N9) avian influenza viruses emerged in China in 2013 and continue to be a threat to human public health,having infected over 800 individuals with a mortality rate approaching 40%. Treatment options for people infected with A(H7N9) include the use of neuraminidase (NA) inhibitors. However,like other influenza viruses,A(H7N9) can become resistant to these drugs. The use of monoclonal antibodies is a rapidly developing strategy for controlling influenza virus infection. Here we generated a murine monoclonal antibody (3c10-3) directed against the NA of A(H7N9) and show that prophylactic systemic administration of 3c10-3 fully protected mice from lethal challenge with wild-type A/Anhui/1/2013 (H7N9). Further,post-infection treatment with a single systemic dose of 3c10-3 at either 24,48 or 72 h post A(H7N9) challenge resulted in both dose- and time-dependent protection of up to 100% of mice,demonstrating therapeutic potential for 3c10-3. Epitope mapping revealed that 3c10-3 binds near the enzyme active site of NA,and functional characterization showed that 3c10-3 inhibits the enzyme activity of NA and restricts the cell-to-cell spread of the virus in cultured cells. Affinity analysis also revealed that 3c10-3 binds equally well to recombinant NA of wild-type A/Anhui/1/2013 and to a variant NA carrying a R289K mutation known to infer NAI resistance. These results suggest that 3c10-3 has the potential to be used as a therapeutic to treat A(H7N9) infections either as an alternative to,or in combination with,current NA antiviral inhibitors.
View Publication
产品号#：
03800
03801
03802
03803
03804
03805
03806
产品名：
ClonaCell™-HY杂交瘤试剂盒
ClonaCell™-HY培养基A
ClonaCell™-HY 培养基 B
ClonaCell™-HY 培养基 C
ClonaCell™-HY 培养基 D
ClonaCell™-HY 培养基 E
ClonaCell™-HY PEG
Yew CW and Tan YJ ( 2016)
1426 225--33
Generation of mouse monoclonal antibodies specific to Chikungunya virus using ClonaCell-HY hybridoma cloning kit
Monoclonal antibodies offer high specificity and this makes it an important tool for molecular biology,biochemistry and medicine. Typically,monoclonal antibodies are generated by fusing mouse spleen cells that have been immunized with the desired antigen with myeloma cells to create immortalized hybridomas. Here,we describe the generation of monoclonal antibodies that are specific to Chikungunya virus using ClonaCell-HY system.
View Publication
产品号#：
03800
03801
03802
03803
03804
03805
03806
产品名：
ClonaCell™-HY杂交瘤试剂盒
ClonaCell™-HY培养基A
ClonaCell™-HY 培养基 B
ClonaCell™-HY 培养基 C
ClonaCell™-HY 培养基 D
ClonaCell™-HY 培养基 E
ClonaCell™-HY PEG
Cindric Vranesic A et al. (MAY 2016)
Human Molecular Genetics May 11 ddw140
Characterization of SKAP/kinastrin isoforms: the N-terminus defines tissue specificity and Pontin binding
Small Kinetochore-Associated Protein (SKAP)/Kinastrin is a multifunctional protein with proposed roles in mitosis,apoptosis and cell migration. Exact mechanisms underlying its activities in these cellular processes are not completely understood. SKAP is predicted to have different isoforms,however,previous studies did not differentiate between them. Since distinct molecular architectures of protein isoforms often influence their localization and functions,this study aimed to examine the expression profile and functional differences between SKAP isoforms in human and mouse. Analyses of various human tissues and cells of different origin by RT-PCR,and by Western blotting and immunocytochemistry applying newly generated anti-SKAP monoclonal antibodies revealed that human SKAP exists in two protein isoforms: ubiquitously expressed SKAP16 and testis/sperm-specific SKAP1. In mouse,SKAP1 expression is detectable in testis at 4 weeks postnatally,when the first wave of spermatogenesis in mice is complete and the elongated spermatids are present in the testes. Furthermore,we identified Pontin as a new SKAP1 interaction partner. SKAP1 and Pontin co-localized in the flagellar region of human sperm suggesting a functional relevance for SKAP1-Pontin interaction in sperm motility. Since most previous studies on SKAP were performed with the testis-specific isoform SKAP1,our findings provide a new basis for future studies on the role of SKAP in both human somatic cells and male germ cells,including studies on male fertility.
View Publication
产品号#：
03800
03801
03802
03803
03804
03805
03806
产品名：
ClonaCell™-HY杂交瘤试剂盒
ClonaCell™-HY培养基A
ClonaCell™-HY 培养基 B
ClonaCell™-HY 培养基 C
ClonaCell™-HY 培养基 D
ClonaCell™-HY 培养基 E
ClonaCell™-HY PEG
Cesaro A et al. (SEP 2012)
PLoS ONE 7 9 e45478
An inflammation loop orchestrated by S100A9 and Calprotectin is critical for development of arthritis
OBJECTIVE: The S100A9 and S100A8 proteins are highly expressed by neutrophils and monocytes and are part of a group of damage-associated molecular pattern molecules that trigger inflammatory responses. Sera and synovial fluids of patients with rheumatoid arthritis (RA) contain high concentrations of S100A8/A9 that correlate with disease activity.backslashnbackslashnMETHODS: In this study,we investigated the importance of S100A9 in RA by using neutralizing antibodies in a murine lipopolysaccharide-synchronized collagen-induced arthritis model. We also used an in vitro model of stimulation of human immune cells to decipher the role played by S100A9 in leukocyte migration and pro-inflammatory cytokine secretion.backslashnbackslashnRESULTS: Treatment with anti-S100A9 antibodies improved the clinical score by 50%,diminished immune cell infiltration,reduced inflammatory cytokines,both in serum and in the joints,and preserved bone/collagen integrity. Stimulation of neutrophils with S100A9 protein led to the enhancement of neutrophil transendothelial migration. S100A9 protein also induced the secretion by monocytes of proinflammatory cytokines like TNFα,IL-1β and IL-6,and of chemokines like MIP-1α and MCP-1.backslashnbackslashnCONCLUSION: The effects of anti-S100A9 treatment are likely direct consequences of inhibiting the S100A9-mediated promotion of neutrophil transmigration and secretion of pro-inflammatory cytokines from monocytes. Collectively,our results show that treatment with anti-S100A9 may inhibit amplification of the immune response and help preserve tissue integrity. Therefore,S100A9 is a promising potential therapeutic target for inflammatory diseases like rheumatoid arthritis for which alternative therapeutic strategies are needed.
View Publication
产品号#：
03800
03801
03802
03803
03804
03805
03806
15028
15068
产品名：
ClonaCell™-HY杂交瘤试剂盒
ClonaCell™-HY培养基A
ClonaCell™-HY 培养基 B
ClonaCell™-HY 培养基 C
ClonaCell™-HY 培养基 D
ClonaCell™-HY 培养基 E
ClonaCell™-HY PEG
RosetteSep™人单核细胞富集抗体混合物
RosetteSep™人单核细胞富集抗体混合物
Flyak AI et al. (FEB 2015)
Cell 160 5 893--903
Mechanism of human antibody-mediated neutralization of Marburg virus
The mechanisms by which neutralizing antibodies inhibit Marburg virus (MARV) are not known. We isolated a panel of neutralizing antibodies from a human MARV survivor that bind to MARV glycoprotein (GP) and compete for binding to a single major antigenic site. Remarkably,several of the antibodies also bind to Ebola virus (EBOV) GP. Single-particle EM structures of antibody-GP complexes reveal that all of the neutralizing antibodies bind to MARV GP at or near the predicted region of the receptor-binding site. The presence of the glycan cap or mucin-like domain blocks binding of neutralizing antibodies to EBOV GP,but not to MARV GP. The data suggest that MARV-neutralizing antibodies inhibit virus by binding to infectious virions at the exposed MARV receptor-binding site,revealing a mechanism of filovirus inhibition.
View Publication
产品号#：
03800
03801
03802
03803
03804
03805
03806
产品名：
ClonaCell™-HY杂交瘤试剂盒
ClonaCell™-HY培养基A
ClonaCell™-HY 培养基 B
ClonaCell™-HY 培养基 C
ClonaCell™-HY 培养基 D
ClonaCell™-HY 培养基 E
ClonaCell™-HY PEG
Fang L et al. (MAY 2008)
The Journal of Experimental Medicine 205 5 1037--48
Essential role of TNF receptor superfamily 25 (TNFRSF25) in the development of allergic lung inflammation
We identify the tumor necrosis factor receptor superfamily 25 (TNFRSF25)/TNFSF15 pair as critical trigger for allergic lung inflammation,which is a cardinal feature of asthma. TNFRSF25 (TNFR25) signals are required to exert T helper cell 2 (Th2) effector function in Th2-polarized CD4 cells and co-stimulate interleukin (IL)-13 production by glycosphingolipid-activated NKT cells. In vivo,antibody blockade of TNFSF15 (TL1A),which is the ligand for TNFR25,inhibits lung inflammation and production of Th2 cytokines such as IL-13,even when administered days after airway antigen exposure. Similarly,blockade of TNFR25 by a dominant-negative (DN) transgene,DN TNFR25,confers resistance to lung inflammation in mice. Allergic lung inflammation-resistant,NKT-deficient mice become susceptible upon adoptive transfer of wild-type NKT cells,but not after transfer of DN TNFR25 transgenic NKT cells. The TNFR25/TL1A pair appears to provide an early signal for Th2 cytokine production in the lung,and therefore may be a drug target in attempts to attenuate lung inflammation in asthmatics.
View Publication
产品号#：
03800
03801
03802
03803
03804
03805
03806
03831
产品名：
ClonaCell™-HY杂交瘤试剂盒
ClonaCell™-HY培养基A
ClonaCell™-HY 培养基 B
ClonaCell™-HY 培养基 C
ClonaCell™-HY 培养基 D
ClonaCell™-HY 培养基 E
ClonaCell™-HY PEG
ClonaCell™-HY 液体 HAT 筛选培养基
Hostetter DR et al. (SEP 2007)
Journal of Biological Chemistry 282 38 27865--74
Hip is a pro-survival substrate of granzyme B
The extended substrate specificity of granzyme B (GrB) was used to identify substrates among the chaperone superfamily. This approach identified Hsp90 and Bag1-L as novel GrB substrates,and an additional GrB cleavage site was identified in the Hsc70/Hsp70-Interacting Protein,Hip. Hsp90,Bag1L,and Hip were validated as GrB substrates in vitro,and mutational analysis confirmed the additional cleavage site in Hip. Because the role of Hip in apoptosis is unknown,its proteolysis by GrB was used as a basis to test whether it has anti-apoptotic activity. Previous work on Hip was limited to in vitro characterization; therefore,it was important to demonstrate Hip cleavage in a physiological context and to show its relevance to natural killer (NK) cell-mediated death. Hip is cleaved at both GrB cleavage sites during NK-mediated cell death in a caspase-independent manner,and its cleavage is due solely to GrB and not other granule components. Furthermore,Hip is not cleaved upon stimulation of the Fas receptor in the Jurkat T-cell line,suggesting that Hip is a substrate unique to GrB. RNA interference-mediated reduction of Hip within the K562 cell line rendered the cells more susceptible to NK cell-mediated lysis,indicating that proteolysis by GrB of Hip contributes to death induction. The small effect of RNA interference-mediated Hip deficiency on cytotoxicity is in agreement with the inherent redundancy of NK cell-mediated cell death. The identification of additional members of the chaperone superfamily as GrB substrates and the validation of Hip as an anti-apoptotic protein contribute to understanding the interplay between stress response and apoptosis.
View Publication
产品号#：
03814
产品名：
ClonaCell™-TCS 培养基
Wittman VP et al. (SEP 2006)
The Journal of Immunology 177 6 4187--95
Antibody targeting to acClass I MHC-peptide epitope promotes tumor cell death
Therapeutic mAbs that target tumor-associated Ags on the surface of malignant cells have proven to be an effective and specific option for the treatment of certain cancers. However,many of these protein markers of carcinogenesis are not expressed on the cells' surface. Instead these tumor-associated Ags are processed into peptides that are presented at the cell surface,in the context of MHC class I molecules,where they become targets for T cells. To tap this vast source of tumor Ags,we generated a murine IgG2a mAb,3.2G1,endowed with TCR-like binding specificity for peptide-HLA-A*0201 (HLA-A2) complex and designated this class of Ab as TCR mimics (TCRm). The 3.2G1 TCRm recognizes the GVL peptide (GVLPALPQV) from human chorionic gonadotropin beta presented by the peptide-HLA-A*0201 complex. When used in immunofluorescent staining reactions using GVL peptide-loaded T2 cells,the 3.2G1 TCRm specifically stained the cells in a peptide and Ab concentration-dependent manner. Staining intensity correlated with the extent of cell lysis by complement-dependent cytotoxicity (CDC),and a peptide concentration-dependent threshold level existed for the CDC reaction. Staining of human tumor lines demonstrated that 3.2G1 TCRm was able to recognize endogenously processed peptide and that the breast cancer cell line MDA-MB-231 highly expressed the target epitope. The 3.2G1 TCRm-mediated CDC and Ab-dependent cellular cytotoxicity of a human breast carcinoma line in vitro and inhibited in vivo tumor implantation and growth in nude mice. These results provide validation for the development of novel TCRm therapeutic reagents that specifically target and kill tumors via recognition and binding to MHC-peptide epitopes.
View Publication
产品号#：
03800
03801
03802
03803
03804
03805
03806
03831
产品名：
ClonaCell™-HY杂交瘤试剂盒
ClonaCell™-HY培养基A
ClonaCell™-HY 培养基 B
ClonaCell™-HY 培养基 C
ClonaCell™-HY 培养基 D
ClonaCell™-HY 培养基 E
ClonaCell™-HY PEG
ClonaCell™-HY 液体 HAT 筛选培养基
Coffman KT et al. (NOV 2003)
Cancer Research 63 22 7907--12
Differential EphA2 epitope display on normal versus malignant cells.
The EphA2 receptor tyrosine kinase is overexpressed in many different types of human cancers where it functions as a powerful oncoprotein. Dramatic changes in the subcellular localization and function of EphA2 have also been linked with cancer,and in particular,unstable cancer cell-cell contacts prevent EphA2 from stably binding its ligand on the surface of adjoining cells. This change is important in light of evidence that ligand binding causes EphA2 to transmit signals that negatively regulate tumor cell growth and invasiveness and also induce EphA2 degradation. On the basis of these properties,we have begun to target EphA2 on tumor cells using agonistic antibodies,which mimic the consequences of ligand binding. In our present study,we show that a subset of agonistic EphA2 antibodies selectively bind epitopes on malignant cells,which are not available on nontransformed epithelial cells. We also show that such epitopes arise from differential cell-cell adhesions and that the stable intercellular junctions of nontransformed epithelial cells occlude the binding site for ligand,as well as this subset of EphA2 antibodies. Finally,we demonstrate that antibody targeting of EphA2 decreases tumor cell growth as measured using xenograft tumor models and found that the mechanism of antibody action relates to EphA2 protein degradation in vivo. Taken together,these results suggest new opportunities for therapeutic targeting of the large number of different cancers that express EphA2 in a manner that could minimize potential toxicities to normal cells.
View Publication
产品号#：
03800
03801
03802
03803
03804
03805
03806
产品名：
ClonaCell™-HY杂交瘤试剂盒
ClonaCell™-HY培养基A
ClonaCell™-HY 培养基 B
ClonaCell™-HY 培养基 C
ClonaCell™-HY 培养基 D
ClonaCell™-HY 培养基 E
ClonaCell™-HY PEG
Sun Y et al. (MAY 2016)
mBio 7 2 e00465--16
Deletion of a Yci1 Domain Protein of Candida albicans Allows Homothallic Mating in MTL Heterozygous Cells
It has been proposed that the ancestral fungus was mating competent and homothallic. However,many mating-competent fungi were initially classified as asexual because their mating capacity was hidden behind layers of regulation. For efficient in vitro mating,the essentially obligate diploid ascomycete pathogen Candida albicans has to change its mating type locus from heterozygous MTL a /α to homozygous MTL a / a or MTL α/α and then undergo an environmentally controlled epigenetic switch to the mating-competent opaque form. These requirements greatly reduce the potential for C. albicans mating. Deletion of the Yci1 domain gene OFR1 bypasses the need for C. albicans cells to change the mating type locus from heterozygous to homozygous prior to switching to the opaque form and mating and allows homothallic mating of MTL heterozygous strains. This bypass is carbon source dependent and does not occur when cells are grown on glucose. Transcriptional profiling of ofr1 mutant cells shows that in addition to regulating cell type and mating circuitry,Ofr1 is needed for proper regulation of histone and chitin biosynthesis gene expression. It appears that OFR1 is a key regulator in C. albicans and functions in part to maintain the cryptic mating phenotype of the pathogen.
View Publication
EasySep™小鼠TIL（CD45）正选试剂盒
沪公网安备31010102008431号