Preparation and identification of monoclonal antibodies against ω-conotoxin MVIIA.
In this study, an MAb against ω-conotoxin MVIIA was prepared. The conotoxin-coding DNA sequence was chemically synthesized and cloned into expression vector pGEX-6p-1 and pET32a (+), respectively. The fusion protein GST-CTX was expressed and purified, and was used to immunize BALB/c mice for preparing the anti-CTX antibody. The spleen cells were fused with SP2/0 myeloma cells after the titer of antiserum was detected and qualified. After being screened by indirect ELISA and cloned by limiting dilution, a hybridoma named 4A12, which produces monoclonal antibody specifically against ω-CTX MVIIA, was successfully ...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Establishment of neutralizing rat monoclonal antibodies for fibroblast growth factor-2.
Authors: Tanaka M, Yamaguchi M, Shiota M, Kawamoto Y, Takahashi K, Inagaki A, Osada-Oka M, Harada A, Wanibuchi H, Izumi Y, Miura K, Iwao H, Ohkawa Y Abstract Fibroblast growth factor-2 (FGF-2) plays a critical role in endothelial survival, proliferation, and angiogenesis and is localized on the cell membrane by binding to heparan sulfate proteoglycans. Here we established a neutralizing monoclonal antibody, 1B9B9, against FGF-2 using the rat medial iliac lymph node method. 1B9B9 blocked the binding of FGF-2 to its receptor, inhibiting FGF-2-induced proliferation and corresponding downstream signaling in endothelial...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Preparation and characterization of monoclonal antibody against human B7-H4 molecule.
In this study, we report on the establishment and characterization of a functional anti-human B7-H4 monoclonal antibody (MAb) 5G3 through hybridoma method. Flow cytometry analysis showed that MAb 5G3 specifically bound to B7-H4 molecule. Functional experiments indicated that MAb 5G3 could block the inhibitory role of B7-H4 molecule on A549 cells in and reduce the apoptosis of Jurkat cells, suggesting that MAb 5G3 is an antagonistic antibody and a useful tool for further studies of B7-H4 functions in cancers. PMID: 25171007 [PubMed - indexed for MEDLINE] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Monoclonal antibody-based immunoassay for analysis of octopamine in housefly.
In this study, a new hapten of OA was synthesized via aldol condensation. With the hapten, monoclonal antibodies (MAb) were generated and their characterizations were investigated. An indirect competitive enzyme-linked immunosorbent assay (icELISA) based on MAb 3C11-E3 was established, which required simple sample pre-treatments and had low cross-reactivity with OA structural analogise. The half maximal inhibition concentration (IC50) and the detected range (IC20-IC80) of the icELISA were 128 ng/mL and 12-1438 ng/mL, respectively. Average recoveries of OA ranged from 73 to 129% in the housefly. PMID: 2517...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

An uncoordinated-5 homolog B receptor monoclonal antibody regulates A375 melanoma cell migration.
In this study, we report on the generation of a monoclonal antibody specific to the outer-membrane immunoglobulin-like domains of UNC5B using the hybridoma technique. Western blot, immunofluorescence, and flow cytometry analyses showed that the antibody specifically bound to UNC5B protein. Interestingly, the antibody blocked the Netrin-1-induced inhibitory effect on the mobility of melanoma A375 cells by wound healing assay and transwell migration assay, whereas it had no effects on cell proliferation measured by CCK-8 assay. Thus, the functional antibody may provide a useful tool for the study of UNC5B expression profiles...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Generation and characterization of polyclonal antibody against part of immunoglobulin constant heavy υ chain of goose.
Generation and characterization of polyclonal antibody against part of immunoglobulin constant heavy υ chain of goose. Monoclon Antib Immunodiagn Immunother. 2014 Aug;33(4):287-90 Authors: Zhao P, Guo Y, Ma B, Xing M, Wang J Abstract Immunoglobulin Y (abbreviated as IgY) is a type of immunoglobulin that is the major antibody in bird, reptile, and lungfish blood. IgY consists of two light (λ) and two heavy (υ) chains. In the present study, polyclonal antibody against IgYFc was generated and evaluated. rIgYCυ3/Cυ4 was expressed in Escherichia coli, purified and util...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Development and characterization of monoclonal antibodies for rapid detection of Acinetobacter baumannii.
Authors: Baig A, Cabral TM, Corbett CR Abstract Recently Acinetobacter baumannii has emerged as a dominant form of nosocomial infections worldwide. As such, diagnostic tools are urgently needed. Reported here is the development/characterization of two mouse monoclonal antibodies (MAbs), F241G3sc2 and F241G6sc2, against A. baumannii ATCC 19606 with clinical strain cross-reactivity. Specifically, both MAbs cross-reacted with 33% of tested A. baumannii clinical strains, without cross-reactivity against other tested species of Acinetobacter. However, further testing with additional clinical strains and species is neede...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

A monoclonal antibody against the extracellular domain of mouse and human epithelial V-like antigen 1 reveals a restricted expression pattern among CD4- CD8- thymocytes.
Authors: Garabatos N, Blanco J, Fandos C, Lopez E, Santamaria P, Ruiz A, Perez-Vidakovics ML, Benveniste P, Galkin O, Zuñiga-Pflucker JC, Serra P Abstract Expression of transcripts for the homotypic adhesion protein epithelial V-like antigen 1 (EVA1), also known as myelin protein zero like-2 (Mpzl2), is known to be present in thymic stromal cells. However, protein expression within different thymic subsets, stromal and/or lymphoid, has not been characterized due a lack of specific reagents. To address this, we generated a hybridoma (G9P3-1) secreting a monoclonal antibody (G9P3-1Mab), reactive against both h...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

A8, an anti-uPA agonistic antibody, promotes metastasis of cancer cells via ERK pathway.
In this study, scFv A8 was converted to minibody form and evaluated for its functional ability on the uPA system involved in cellular signaling and cancer cell metastasis. A8 minibody increased enzyme activity of uPA and enhanced the migration and invasion of HT1080 colon cancer cells in a dose-dependent manner. A8 increased ERK phosphorylation, and enhanced migration was blocked by U0126, but not by LY0294002, SB2203580, and SP600125. A8 minibody also enhanced migration of MDA-MB231 by mediated expressing surface uPA, but not that of MCF-7 non-expressing surface uPA. Taken together, the A8 anti-uPA antibody is a uPA agoni...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Development and characterization of three novel monoclonal antibodies against CA-125.
We describe here the generation and characterization of three novel hybridoma cell lines producing MAbs against CA-125. CA-125 purified from culture supernatant of ovarian carcinoma cell line OVCAR-3 by affinity chromatography, was used for immunization of BALB/c mice. Three stable cell lines (3C8, 2B6, and 5A12) were selected for production of antibodies against CA-125 and were expanded in mass culture. All three antibodies were shown to recognize linear epitopes. Antibodies 2B6 and 5A12 were determined to recognize epitope cluster B (M 11-like); MAb 3C8 was classified as group A-epitope binders (OC 125-like). The antibod...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Investigation of hFVIII production in mammary glands of transgenic mice.
Authors: Mohammadian T, Rassi H Abstract Hemophilia A is an X-linked disorder affecting 1 in 10,000 males. The disease is caused by a defect or mutation of factor 8 or 9. Human factor 8 gene (hFVIII) is a relatively large gene consisting of 26 exons and approximately 2,351 amino acids with a length of 9 Kb mRNA. Expression of hFVIII in mammalian milk is becoming a widespread strategy for high-level production of hFVIII because of the most complex post-translational modifications. The aim of this study was the cloning and expression of hFVIII in mammary glands of two transgenic mice. To obtain a recombinant plasmid,...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Development and application of an ELISA kit for the detection of milk progesterone in dairy cows.
Authors: Wu L, Xu C, Xia C, Duan Y, Xu C, Zhang H, Bao J Abstract Progesterone (P4) is a steroid gonadal hormone that is mainly produced from the corpus luteum and placenta and has various biological functions, especially reproductive regulation. It is important to establish a specific and sensitive P4 enzyme-linked immunosorbent assay (ELISA) for the study of ovary activity and reproductive disorders. Therefore, we prepared a monoclonal antibody (MAb) in a completed antigen (11α-OH-P4-HS-OVA). Based on the MAb and our previously prepared completed antigen, a highly specific and sensitive ELISA was developed....
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Development of monoclonal antibody-based sandwich ELISA for detection of dextran.
Authors: Wang SY, Li Z, Wang XJ, Lv S, Yang Y, Zeng LQ, Luo FH, Yan JH, Liang DF Abstract Dextran as anti-nutritional factor is usually a result of bacteria activity and has associated serial problems during the process stream in the sugar industry and in medical therapy. A sensitive method is expected to detect dextran quantitatively. Here we generated four monoclonal antibodies (MAbs) against dextran using dextran T40 conjugated with bovine serum albumin (BSA) as immunogen in our lab following hybridoma protocol. Through pairwise, an MAb named D24 was determined to be conjugated with horseradish peroxidase (HRP) ...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Monoclonal antibodies against G protein of spring viremia of carp virus.
Authors: Luo P, Ruan X, Zhang Q, Li Z, Wang M, Liu X Abstract Part of the G protein (3094-4170 bp) of spring viremia of carp virus (SVCV) was expressed in Escherichia coli and purified by dialysis in our study. Two clones of monoclonal antibodies (MAbs 1H11 and 4B8) against G protein were generated by fusion of mouse myeloma cell line SP2/0 and spleen lymphocytes from part of G protein (3094-4170 bp) immunized mice. The results of ELISA (enzyme-linked immunosorbent assay), IFA (indirect immunofluorescent assay), and Western blot assay further demonstrated the characterizations of the two MAbs. Both 1H11 and 4B8 wer...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Development of monoclonal antibodies specifically recognizing the endogenous sterile alpha motif and HD domain 1 protein in porcine cell lines.
In conclusion, specific MAbs of porcine SAMHD1 are reported, and these MAbs provide a valuable tool for further studies of SAMHD1-mediated signaling in virus-infected cells to elucidate the underlying antiviral mechanism. PMID: 25358004 [PubMed - indexed for MEDLINE] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Preparation of monoclonal antibodies against bovine haptoglobin.
Authors: Wang C, Gu C, Guo D, Gao J, Li C, Liu N, Geng Y, Su M, Wang X, Sun D Abstract Female, 8-week-old BALB/c mice were immunized with purified recombinant proteins of the predicted immunodominant region of bovine haptoglobin (pirBoHp). Two monoclonal antibodies (MAbs), named 1B3 and 6D6, were prepared by conventional B lymphocyte hybridoma technique. Titers of ascitic fluid and cell culture supernatant of MAb 1B3 were 1:9.6 × 10(8) and 1:8.2 × 10(4), respectively, and that of MAb 6D6 were 1:4.4 × 10(5) and 1:1.0 × 10(4), respectively. The subtype of MAbs 1B3 and 6D6 was IgG1κ. In W...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Generation of a monoclonal antibody recognizing mouse PD-1.
Authors: Qin YE, Wang G, Yang J, Liu CQ Abstract Programmed death-1 (PD-1) is a transmembrane protein that shares homology with the B7/CD28 family of T cell signaling molecules. PD-1 interacts with its ligands PD-L1 and/or PD-L2 and provides a negative regulatory signal to CD4 and CD8 T cells that results ultimately in a phenotype termed T cell exhaustion. Here we expressed and purified mouse PD-1 protein and developed a monoclonal antibody (MAb) against mouse PD-1 by immunizing BALB/c mice with a specific region of the extracellular domains of PD-1 as antigen, which was expressed in Escherichia coli. A stable hybr...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Production of a Chaetomium globosum enolase monoclonal antibody.
In this study, we describe the production and initial characterization of a monoclonal antibody (MAb) for C. globosum enolase. MAb 1C7, a murine IgG1 isotype MAb, was produced and reacted with recombinant C. globosum enolase (rCgEno) in an enzyme-linked immunosorbent assay and with a putative C. globosum enolase in a Western blot. Epitope mapping showed MAb 1C7 specific reactivity to an enolase decapeptide, LTYEELANLY, that is highly conserved within the fungal class Sordariomycetes. Cross-reactivity studies showed MAb 1C7 reactivity to C. atrobrunneum but not C. indicum. MAb 1C7 did not react with enolase from Aspergillus...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Development and characterization of monoclonal antibodies to detect klotho.
Authors: Maltare A, Nietz AK, Laszczyk AM, Dunn TS, Ballestas ME, Accavitti-Loper MA, King GD Abstract Although antibodies are commercially available to allow investigation into the biology of the age-regulating protein Klotho, problems with antibody specificity and application functionality are significant barriers to progress. Chief among these limitations is the inability of current tools to allow in vivo validation of binding partners originally identified through transfection of tagged proteins. To overcome this barrier, we generated a series of hybridoma cell lines by immunizing rats with a GST-KL1 fusion pro...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Generation and characterization of a monoclonal antibody against prM protein of West Nile virus.
In this study we generated and characterized a monoclonal antibody (MAb) against the WNV prM protein. Western blot analysis showed that the MAb reacted with WNV prM specifically. Immunohistochemistry assays demonstrated that the MAb recognized native prM protein in transfected BHK-21 cells. Preliminary studies were performed to identify the epitope recognized by the MAb using a set of synthesized overlapping peptides spanning the whole length of the prM protein. The MAb reported here may provide a valuable tool for the further exploration of the biological properties and functions of the prM protein and may also be develop...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Cloning murine antibody V-genes with non-degenerate primers and conversion to a recombinant antibody format.
Authors: Bialon M, Schellenberg L, Herzog N, Kraus S, Jörißen H, Fischer R, Stein C, Nähring J, Barth S, Püttmann C Abstract Monoclonal antibodies are produced in cultured hybridoma cell lines, but these cells tend to be unstable; it is therefore necessary to rescue the corresponding genetic information. Here we describe an improved method for the amplification of antibody variable gene (V-gene) information from murine hybridoma cells using a panel of specific, non-degenerate primers. This primer set allows sequences to be rescued from all murine V-genes, except the lambda light chain genes, wh...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Proteoliposome-based selection of a recombinant antibody fragment against the human M2 muscarinic acetylcholine receptor.
Authors: Suharni, Nomura Y, Arakawa T, Hino T, Abe H, Nakada-Nakura Y, Sato Y, Iwanari H, Shiroishi M, Asada H, Shimamura T, Murata T, Kobayashi T, Hamakubo T, Iwata S, Nomura N Abstract The development of antibodies against human G-protein-coupled receptors (GPCRs) has achieved limited success, which has mainly been attributed to their low stability in a detergent-solubilized state. We herein describe a method that can generally be applied to the selection of phage display libraries with human GPCRs reconstituted in liposomes. A key feature of this approach is the production of biotinylated proteoliposomes that ca...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Detection of constant domain of human T cell antigen receptor alpha-chain via novel monoclonal antibody 7F18.
Authors: Reed BK, Lee KA, Bell MP, Gil D, Schrum AG Abstract The αβ T cell antigen receptor (TCR) endows T lymphocytes with immune specificity and controls their effector functions. Each person possesses a vast repertoire of TCRs that is generated by the well-studied processes of somatic recombination and thymic selection. While many antibodies specific for TCRβ variable domains are available, antibodies specific for human TCRα are rare. We now report a novel monoclonal antibody, 7F18, which binds to human TCRα constant region, with specificity for a denatured epitope that can be visuali...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Monoclonal antibodies specific for human IgM Fc receptor inhibit ligand-binding activity.
Authors: Kubagawa Y, Honjo K, Kang DW, Kubagawa H Abstract A panel of six different murine hybridoma clones secreting IgG monoclonal antibodies (MAbs) specific for the human IgM Fc receptor (FcμR) was generated. All MAbs specifically precipitated a major protein of ∼60 kDa from membrane lysates of FcμR-bearing, but not FcμR-negative, cells as did IgM-ligands. Pre-incubation of membrane lysate of FcμR-bearing cells with these MAbs completely removed the ∼60 kDa IgM-reactive protein. By using recombinant human/mouse chimeric FcμR proteins, the epitope recognized by HM7 and HM10 MA...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Characterization of a monoclonal antibody against CREPT, a novel protein highly expressed in tumors.
This study provided a basis for large-scale production of the CREPT antibody, which will be useful for the study of the role of CREPT in different tumors. PMID: 25545209 [PubMed - in process] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Production of monoclonal antibody against EP0 protein of pseudorabies virus and determination of its recognized epitope.
In this study, six monoclonal antibodies (MAbs) against EP0 protein of PRV were generated and their characterizations were investigated. Western blot analysis showed all six MAbs could react with immunizing antigen, but only 2B12 and 2C6 could react with native EP0 protein from PRV-infected cells. ELISA additivity tests revealed that at least three epitopes in EP0 were defined by six MAbs. The epitope recognized by MAb 2B12 was further identified in 287-292 aa of EP0 protein using a series of expressed overlapping peptides. These MAbs may provide valuable tools for further research of the functions of EP0 in PRV infection....
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Production and characterization of highly specific monoclonal antibodies to D-glutamic acid.
In this study, a highly specific monoclonal antibody to D-glutamic acid (D-Glu-MAb) was produced using a hybridoma method. Characterization of D-Glu-MAb by indirect enzyme-linked immunosorbent assay (ELISA) revealed that it has high selectivity against D-Glu-glutaraldehyde (GA) conjugates, while no cross-reaction was observed when 38 other kinds of AA-GA conjugates were used. Moreover, subsequent indirect competitive ELISA disclosed that an epitope of D-Glu-MAb is a D-Glu-GA molecule in the conjugates, suggesting that D-Glu-MAb could be a useful tool to investigate the functional analysis of D-Glu in immunostaining. P...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Preparation of monoclonal antibodies against mycobacterium tuberculosis TB10.4 antigen.
In this study, the gene encoding TB10.4 was cloned into prokaryotic expression vecters pET-30(a) and pGEX-6p-1. The two recombinant proteins His-TB10.4 and GST-TB10.4 were then expressed in vitro in prokaryotic expression systems to develop monoclonal antibodies (MAbs) against TB10.4 protein. The purified rHis-TB10.4 protein was used to immunize BALB/c mice, and eight MAbs were produced. An immunoblotting analysis indicated that all these MAbs specifically recognize the TB10.4 protein. These new MAbs provide powerful reagents for further functional research into TB10.4 protein. PMID: 25545212 [PubMed - in process] (So...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Festschrift dedicated to Hilary Koprowski.
Authors: PMID: 25682617 [PubMed - indexed for MEDLINE] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Anti-mouse properdin TSR 5/6 monoclonal antibodies block complement alternative pathway-dependent pathogenesis.
In this report we use a portion of the mouse properdin protein, expressed in a bacterial system, to raise rabbit polyclonal and hamster monoclonal antibodies that block properdin-dependent pathogenesis. These antibodies, when employed with AP-dependent mouse disease models, can help evaluate the feasibility of properdin-directed therapy. PMID: 25723276 [PubMed - indexed for MEDLINE] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

A novel marker for undifferentiated human embryonic stem cells.
Authors: Higashi K, Yagi M, Arakawa T, Asano K, Kobayashi K, Tachibana T, Saito K Abstract Human embryonic stem cells (hESCs) are pluripotent stem cells from early embryos, and their self-renewal capacity depends on the sustained expression of hESC-specific molecules and the suppressed expression of differentiation-associated genes. To discover novel molecules expressed on hESCs, we generated a panel of monoclonal antibodies against undifferentiated hESCs. The antigen recognized by MAb2 is expressed on the cell surface of undifferentiated hESCs; three diffused bands with molecular mass between 30 and 60 kDa ...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Development of a hybridoma cell line secreting monoclonal antibody against S protein of a Chinese variant of PEDV.
Authors: Lei X, Zhao J, Wang X, Zhao Y, Wang C Abstract Gene encoding 22-380 aa of spike protein of porcine epidemic diarrhea virus (PEDV) isolate in China was cloned and expressed as a recombinant protein in Escherichia coli BL21(DE3). Female BALB/c mice were immunized with the purified recombinant S protein, and a monoclonal antibody (MAb) designated as 2D1 against S protein was achieved by hybridoma technique. MAb 2D1 reacted with S protein of PEDV specifically. The monoclonal antibody 2D1 may provide a useful tool as a specific diagnostic reagent for detecting S protein of the Chinese variant of PEDV and for fu...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Development and characterization of neutralizing monoclonal antibodies against the S1 subunit protein of QX-like avian infectious bronchitis virus strain Sczy3.
Authors: Zou N, Wang F, Duan Z, Xia J, Wen X, Yan Q, Liu P, Cao S, Huang Y Abstract Infectious bronchitis (IB) is a highly contagious disease in chickens caused by infectious bronchitis virus (IBV). The present study was carried out with the aim to develop anti-spike 1 (S1) subunit monoclonal antibodies (MAbs) that could react with IBV strains of different genotypes. The high antigenicity region of S1 gene of an QX-like IBV strain Sczy3 was amplified and ligated into the prokaryotic expression vector pET-32a(+), and the recombinant His-S1 fusion proteins were expressed and purified. The purified whole viral antigen...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

A novel monoclonal antibody against human GRK6 antigen.
In this study, a 20-aa-long peptide of human GRK6 C-terminus domain was synthesized and covalently coupled to keyhole limpet hemocyanin (KLH). A mouse monoclonal antibody against human GRK6 (anti-GRK6 MAb) was successfully prepared through hybridoma technique by immunizing BALB/c mice with synthesized GRK6426-446-KLH peptides. A high specificity and affinity strain of hybridoma 5D12 were established. The titer of the purified anti-GRK6 MAb was 1.28 × 10(6) measured by indirect ELISA. Western blot and immunocytochemistry experiments were also applied to characterize the antibody specificity. Antibody absorption assays...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Generation of med28 specific monoclonal antibodies.
In this study, we report for the first time the generation and characterization of four monoclonal antibodies against med28 through immunoblotting, immunofluorescence microscopy, immunoprecipitation, and immunohistochemical analyses. These antibodies will be useful in detecting med28 in in vitro and in vivo. PMID: 25723281 [PubMed - indexed for MEDLINE] (Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy)
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Production and characterization of a peptide-based monoclonal antibody against CD44 variant 6.
In this study, a synthetic peptide from CD44v6 was conjugated to keyhole limpet hemocyanin (KLH) and injected into BALB/c mice. Splenocytes from the immunized mice were fused with murine SP2/0 myeloma cells followed by selection of antibody producing hybridoma cells. After screening of hybridoma colonies by ELISA, high affinity antibodies were selected and purified by affinity chromatography. Western blot, immunocytochemistry, and flow cytometry experiments were used to characterize the antibodies. Six stable hybridoma cell lines, designated as 1H1, 1H2, 2A12, 2G11, 3H3, and 3H7, were obtained. Flow cytometry and immunocyt...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Characterization of monoclonal antibody LpMab-3 recognizing sialylated glycopeptide of podoplanin.
In this study, we produced and characterized a novel anti-podoplanin monoclonal antibody, LpMab-3, the epitope of which is a sialylated glycopeptide of podoplanin. We identified the minimum epitope of LpMab-3 as Thr76-Glu81 of human podoplanin, which is different from PLAG domain, using Western blot analysis and flow cytometry. Immunohistochemical analysis showed that LpMab-3 is useful for detecting lung type I alveolar cells and lymphatic endothelial cells. Because LpMab-3 detects only sialylated podoplanin, it could be useful for uncovering the physiological function of sialylated human podoplanin. PMID: 25723283 [P...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Monoclonal antibody to N protein of porcine epidemic diarrhea virus.
Authors: Pan X, Kong N, Shan T, Zheng H, Tong W, Yang S, Li G, Zhou E, Tong G Abstract The N gene of porcine epidemic diarrhea virus (PEDV) was amplified by RT-PCR using specific primers, and inserted into the expression vector pCold-I to construct a recombinant plasmid pCold-I-N. The recombinant plasmid was expressed in Escherichia coli BL21 (DE3) under IPTG induction. Then, female BALB/c mice were immunized with the purified recombinant N protein and one strain of hybridoma cells named 2B8 secreting anti-N protein monoclonal antibodies (MAb) was obtained by hybridoma technique. The MAb was specifically reacted wi...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

New monoclonal antibodies specific for 1-(5-fluoropentyl)-3-(2-iodobenzoyl)indole.
Authors: Nakayama H, Kenjyou N Abstract 1-(5-fluoropentyl)-3-(2-iodobenzoyl)indole (AM694) is one of the synthetic cannabinoids and an illegal drug in Japan. It is important to generate a monoclonal antibody (MAb) against AM694 for use in the rapid and sensitive detection of the drug. Two monoclonal antibodies, named HN0124 (IgG1) and NK0504 (IgG1), were obtained, which were possibly effective for detecting AM694 and its derivatives. The cross-reactive ability of these MAbs was evaluated using a competitive enzyme-linked immunosorbent assay. In the results, both of these antibodies recognize 1-(5-fluoropentyl)-3-(2...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Functional and Molecular Characteristics of Novel and Conserved Cross-Clade HIV Envelope Specific Human Monoclonal Antibodies.
Authors: Kobie JJ, Zheng B, Piepenbrink MS, Hessell AJ, Haigwood NL, Keefer MC, Sanz I Abstract To define features of the B cell response to HIV that may be translated to vaccine development, we have isolated a panel of monoclonal antibodies (MAbs) from HIV-infected patients. These MAbs are all highly reactive to HIV envelope (Env) from multiple clades, and include gp120 and gp41 specificities. Three of the MAbs exhibit substantial homology to previously described VH1-69, VH3-30, and VH4-59 HIV broadly neutralizing antibody lineages. An inherently autoreactive VH4-34 encoded MAb was reactive to diverse Env despite ...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Defining Potential Vaccine Targets of Haemophilus ducreyi Trimeric Autotransporter Adhesin DsrA.
Authors: Fusco WG, Choudhary NR, Stewart SM, Alam SM, Sempowski GD, Elkins C, Leduc I Abstract Haemophilus ducreyi is the causative agent of the sexually transmitted genital ulcer disease chancroid. Strains of H. ducreyi are grouped in two classes (I and II) based on genotypic and phenotypic differences, including those found in DsrA, an outer membrane protein belonging to the family of multifunctional trimeric autotransporter adhesins. DsrA is a key serum resistance factor of H. ducreyi that prevents binding of natural IgM at the bacterial surface and functions as an adhesin to fibronectin, fibrinogen, vitronectin...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Therapeutic Potential of Monoclonal IgA Antibodies in Allergic Diseases: Suppressive Effect of IgA on Immune Responses Induced By Re-exposure to Antigen in Sensitized Mice by Monoclonal IgE Antibody That Binds to a Different Epitope of the Same Antigen.
In this study, we investigated the therapeutic potential of allergen-specific monoclonal IgA on allergic diseases. This antibody acts by decreasing immune responses upon exposure to allergens in mice previously sensitized by a monoclonal IgE that recognizes the allergen. The lack of inhibitory effects of anti-ovalbumin monoclonal IgA (OA-4) on either the binding of anti-ovalbumin monoclonal IgE (OE-1) to ovalbumin by ELISA or on ovalbumin-induced degranulation of rat basophilic leukemia RBL2H3 cells sensitized with OE-1 indicated that OA-4 and OE-1 recognized different epitopes on ovalbumin. Immune responses (anti-ovalbumi...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Monoclonal antibodies against hepatitis B viral surface antigens and epitope grouping.
Authors: Lee G, Liu S Abstract Monoclonal antibodies (MAbs) were generated against subtypes (ad/ad/rw) of the human hepatitis B viral surface antigen (HBsAg). Among dozens of antibodies that were generated, the majority was shown to commonly react with various ad/ay subtypes of the S protein. Epitope(s) of these antibodies were grouped by various immunoassay methods, and at least four distinct epitope regions were identified. Some of these antibodies were selected to formulate sandwich enzyme immunoassays for quantitative determinations of HBsAg in reconstituted specimens. Epitope-defined monoclonal antibodies with...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Monoclonal Antibodies Against NS2B of Japanese Encephalitis Virus.
Authors: Dong Q, Xu Q, Ruan X, Huang S, Cao S Abstract Japanese encephalitis (JE) is one of the most important viral encephalitis, caused by the Japanese encephalitis virus (JEV). The function of non-structural protein 2B (NS2B) mostly remains unclear. In our study, NS2B of Japanese encephalitis virus (JEV) was expressed in Escherichia coli and purified by dialysis. After fusing mouse myeloma cell line SP2/0 with spleen lymphocytes from NS2B protein immunized mice, three clones of monoclonal antibodies (MAbs), named 1B9, 3E12, and 4E6, were generated. The specificity and sensitivity of MAbs were demonstrated by ELI...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Monoclonal Antibodies Against the Human Respiratory Syncytial Virus Obtained by Immunization with Epitope Peptides and CpG-DNA-liposome Complex.
Authors: Park BK, Choi SH, Kim YE, Park S, Lee Y, Lee KW, Kwon HJ Abstract Respiratory syncytial virus (RSV) is the major cause of pulmonary inflammation in infants, young children, and immunocompromised adults. However, the RSV vaccine is not yet available commercially. The RSV-F glycoprotein mediates virus-host cell fusion, leading to syncytial formation; therefore, the RSV-F glycoprotein has been a treatment target for prevention and therapy of RSV infection. To produce the RSV-F-protein epitope-specific monoclonal antibody (MAb), BALB/c mice were immunized with a complex consisting of epitope peptide and MB-ODN...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Generation and Characterization of a New Monoclonal Antibody Against CXCL4.
In this study, we aimed to develop a neutralizing antibody against both human and mouse CXCL4. Rats were immunized with recombinant human CXCL4 (rhCXCL4). Hybridoma clones were created by fusion of the immunized rat spleen cells with mouse myeloma SP2/0 cells and screened using recombinant mouse CXCL4 (rmCXCL4) and rhCXCL4. The CXCL4 monoclonal antibody (CXCL4 MAb) produced by the 16D6-3 hybridoma clone was sequenced and characterized by Western blot and Biacore assays. It recognized both human and mouse CXCL4 with high affinity and neutralized the effect of rhCXCL4 in vitro. Thus, the antibody may be used in the studies o...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Preparation and Characterization of a Monoclonal Antibody Against the Core Protein VP7 of the 25th Serotype of Bluetongue Virus.
Authors: Wu X, Liu Q, He J, Zang M, Wang H, Li Y, Tang L Abstract Bluetongue virus (BTV) is a member of the genus Orbivirus, within the family Reoviridae. The VP7 protein of BTV is used for developing group-specific serological assays. To prepare monoclonal antibody (MAb) against VP7 of the 25th serotype BTV, the RNA S7 encoding VP7 was cloned into prokaryotic expression vectors pET-28a (+) and pGEX-6P-1 to generate recombinant plasmids. The recombinant protein VP7 was expressed in Escherichia coli BL21 (DE3), respectively. The results of SDS-PAGE revealed that the VP7 was expressed and the molecular mass of recomb...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Monoclonal antibody against M protein of spring viremia of carp virus.
Authors: Li Z, Zhang Q, Luo P, Liu G, Wang M, Liu X Abstract M protein of spring viremia of carp virus (SVCV) was expressed in Escherichia coli and then used to immunize BALB/c mice. One monoclonal antibody (5A1) against M protein was generated by fusion of mouse myeloma cell line SP2/0 and spleen lymphocytes from immunized mice. The characterizations of this MAb were confirmed by ELISA (enzyme linked immunosorbent assay), IFA (immunofluorescent assay), and Western blot analysis. All results indicate that MAb 5A1 was specific to SVCV M protein. PMID: 25897611 [PubMed - in process] (Source: Monoclonal Antibodie...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

Camel Heavy Chain Polyclonal Antibody Raised Against Recombinant Murine Placental Growth Factor Expressed in Escherichia coli.
In this study, murine PlGF was cloned, expressed in an Escherichia coli system, and used for development of polyclonal camel antibody. Codon-optimized mouse PlGF (mPlGF) cDNA was synthesized and cloned in to pET-28a. The expression was performed in BL21 (DE3) E. coli strain and purified by immobilized metal affinity column chromatography. A camel was subcutaneously immunized six times over a one-week interval using purified protein. IgGs were purified by applying the serum on two sequential column affinity chromatography using proteins A and G. Then, anti-PlGF IgG was identified by Western bolt analysis and ELISA using com...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research

The Monospecificity of Novel Anti-HLA-E Monoclonal Antibodies Enables Reliable Immunodiagnosis, Immunomodulation of HLA-E, and Upregulation of CD8+ T Lymphocytes.
Authors: Ravindranath MH, Terasaki PI, Pham T, Jucaud V Abstract In human cancers, over-expression of HLA-E is marked by gene expression. However, immunolocalization of HLA-E on tumor cells is impeded by the HLA-Ia reactivity of commercial anti-HLA-E monoclonal antibodies (MAbs). So there was a clear need to develop monospecific anti-HLA-E MAbs for reliable immunodiagnosis of HLA-E, particularly considering the prognostic relevance of HLA-E in human cancer. HLA-E overexpression is correlated with disease progression and poor survival of patients, both of which are attributed to the suppression of anti-tumor activit...
Source: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy - December 2, 2015 Category: Microbiology Tags: Monoclon Antib Immunodiagn Immunother Source Type: research