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https://www.readbyqxmd.com/read/28531814/targeting-egfr-her2-heterodimerization-with-a-novel-anti-her2-domain-ii-iii-antibody
#1
Xiaojie Yu, Lingfei Wang, Yafeng Shen, Chao Wang, Yajun Zhang, Yanchun Meng, Yang Yang, Beibei Liang, Bo Zhou, Huajing Wang, Huafeng Wei, Changhai Lei, Shi Hu, Bohua Li
HER2, a ligand-free tyrosine kinase receptor of the HER family, is frequently overexpressed in breast cancer. The anti-HER2 antibody trastuzumab has shown significant clinical benefits in metastatic breast cancer. Despite the effectiveness of trastuzumab, its efficacy remains variable and often modest. Thus, there is an urgent need to improve ErbB2-targeting therapy. Here, we describe a novel anti-HER2 antibody, 7C3, which was developed using hybridoma technique. Structural analysis confirms that the epitope of this antibody is in domain II/III of HER2...
May 19, 2017: Molecular Immunology
https://www.readbyqxmd.com/read/28529510/advances-in-the-isolation-of-specific-monoclonal-rabbit-antibodies
#2
REVIEW
Zaibao Zhang, Huijuan Liu, Qian Guan, Lei Wang, Hongyu Yuan
The rabbit monoclonal antibodies (mAbs) have advantages in pharmaceuticals and diagnostics with high affinity and specificity. During the past decade, many techniques have been developed for isolating rabbit mAbs, including single B cell antibody technologies. This review describes the basic characterization of rabbit antibody repertoire and summarizes methods of hybridoma technologies, phage display platform, and single B cell antibody technologies. With advances in antibody function and repertoire analysis, rabbit mAbs will be widely used in therapeutic applications in the coming years...
2017: Frontiers in Immunology
https://www.readbyqxmd.com/read/28522882/monoclonal-suncus-antibodies-generation-of-fusion-partners-to-produce-suncus-suncus-hybridomas
#3
Yoshikazu Sado, Satoko Inoue, Yasuko Tomono, Makoto Matsuyama, Masaki Fukushima, Toshitaka Oohashi, Takamichi Jogahara, Sen-Ichi Oda
We used suncus (Suncus murinus; house musk shrew) to generate partner cells for cell fusion to produce suncus monoclonal antibodies. Suncus are insectivores that are genetically distant to rodents, and recognize antigens and epitopes that are not immunogenic in mice and rats, which are the animals most commonly used in basic life science research and from which monoclonal antibodies are usually produced. To date, monoclonal antibodies from suncus have not been generated due to the lack of a plasmacytoma fusion partner...
April 27, 2017: Acta Histochemica et Cytochemica
https://www.readbyqxmd.com/read/28512452/towards-identifying-protective-b-cell-epitopes-the-pspa-story
#4
REVIEW
Naeem Khan, Arif T Jan
Pneumococcal surface protein A (PspA) is one of the most abundant cell surface protein of Streptococcus pneumoniae (S. pneumoniae). PspA variants are structurally and serologically diverse and help evade complement-mediated phagocytosis of S. pneumoniae, which is essential for its survival in the host. PspA is currently been screened for employment in the generation of more effective (serotype independent) vaccine to overcome the limitations of polysaccharide based vaccines, providing serotype specific immune responses...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28505517/a-modified-adenovirus-vector-mediated-antibody-screening-method-identifies-epha2-as-a-cancer-target
#5
Toshihiro Tanaka, Hiromi Yamada, Masahide Kuroki, Shohta Kodama, Kazuo Tamura, Yasushi Takamatsu
BACKGROUND: We constructed a genetically modified adenovirus vector incorporating an IgG Fc-binding motif from staphylococcal protein A, Z33 (Adv-FZ33). Adv-FZ33 allows an antibody to redirect the vector to a target molecule on the cell surface. We attempted to search for target antigen candidates and antibodies that allowed highly selective gene transduction into malignant tumors. METHODS: Hybridoma libraries producing monoclonal antibodies (mAbs) were screened that increased transduction efficiency in cancer cell lines after cross-linking with Adv-FZ33...
May 12, 2017: Translational Oncology
https://www.readbyqxmd.com/read/28502308/-preparation-and-identification-of-monoclonal-antibody-against-h1n1-influenza-virus
#6
Yang Liu, Xiangrong Zhao, Chunyan Guo, Yan Li, Guanghua Wang, Daoyan Liang, Pengbo Yu, Yuan Li, Jun Hu
Objective To prepare the monoclonal antibody (mAb) against influenza A virus (H1N1) using purified viral particles as antigen and investigate the characterization of host cells infected with influenza virus utilizing the mAb. Methods A/PR/8 (H1N1) virus was cultured in embryonated chicken eggs and further purified by differential and density gradient centrifugation. The structure of viral particles was identified by transmission electron microcopy (TEM). Immunogenicity of purified virus was evaluated by Balb/c mice immunized with formalin-inactivated virus...
May 2017: Xi Bao Yu Fen Zi Mian Yi Xue za Zhi, Chinese Journal of Cellular and Molecular Immunology
https://www.readbyqxmd.com/read/28499450/intrabodies-against-the-polysialyltransferases-st8siaii-and-st8siaiv-inhibit-polysialylation-of-ncam-in-rhabdomyosarcoma-tumor-cells
#7
Stefan Somplatzki, Martina Mühlenhoff, Andrea Kröger, Rita Gerardy-Schahn, Thomas Böldicke
BACKGROUND: Polysialic acid (polySia) is a carbohydrate modification of the neural cell adhesion molecule (NCAM), which is implicated in neural differentiation and plays an important role in tumor development and metastasis. Polysialylation of NCAM is mediated by two Golgi-resident polysialyltransferases (polyST) ST8SiaII and ST8SiaIV. Intracellular antibodies (intrabodies; IB) expressed inside the ER and retaining proteins passing the ER such as cell surface receptors or secretory proteins provide an efficient means of protein knockdown...
May 12, 2017: BMC Biotechnology
https://www.readbyqxmd.com/read/28479401/development-of-monoclonal-antibodies-against-igm-of-half-smooth-tongue-sole-cynoglossus-semilaevis-and-analysis-of-phagocytosis-of-fluorescence-microspheres-by-migm-lymphocytes
#8
Shun Yang, Xiaoqian Tang, Xiuzhen Sheng, Jing Xing, Wenbin Zhan
In this work, the two positive hybridomas designated as 1B2 and 2G9 secreting monoclonal antibodies (MAbs) against half-smooth tongue sole IgM were produced and then cloned by limited dilution. Western blotting showed that MAbs 1B2 and 2G9 could specifically react with the heavy and light chains of IgM, respectively. The indirect immunofluorescence assay (IFA) showed that green fluorescent signals were detected on the membrane of lymphocytes, which indicated that membrane-bound IgM (mIgM) could be recognized by both MAbs 1B2 and 2G9...
May 4, 2017: Fish & Shellfish Immunology
https://www.readbyqxmd.com/read/28472993/humanized-monoclonal-antibody-armanezumab-specific-to-n-terminus-of-pathological-tau-characterization-and-therapeutic-potency
#9
Michael G Agadjanyan, Karen Zagorski, Irina Petrushina, Hayk Davtyan, Konstantin Kazarian, Maxim Antonenko, Joy Davis, Charles Bon, Mathew Blurton-Jones, David H Cribbs, Anahit Ghochikyan
BACKGROUND: The experience from clinical trials indicates that anti-Aβ immunotherapy could be effective in early/pre-clinical stages of AD, whereas at the late stages promoting the clearing of Aβ alone may be insufficient to halt the disease progression. At the same time, pathological tau correlates much better with the degree of dementia than Aβ deposition. Therefore, targeting pathological tau may provide a more promising approach for the treatment of advanced stages of AD. Recent data demonstrates that the N-terminal region of tau spanning aa 2-18 termed "phosphatase activation domain" that is normally hidden in the native protein in 'paperclip'-like conformation, becomes exposed in pathological tau and plays an essential role in the inhibition of fast axonal transport and in aggregation of tau...
May 5, 2017: Molecular Neurodegeneration
https://www.readbyqxmd.com/read/28463837/adoptive-cellular-gene-therapy-for-the-treatment-of-experimental-autoimmune-polychondritis-ear-disease
#10
Bin Zhou, Yonggan Liao, Yunkai Guo, Ingo H Tarner, Chunfen Liao, Sisi Chen, Mohammad Habiby Kermany, Hanjun Tu, Sen Zhong, Peijie Chen
In the past, the clinical therapy for autoimmune diseases, such as autoimmune polychondritis ear disease, was mostly limited to nonspecific immunosuppressive agents, which could lead to variable responses. Currently, gene therapy aims at achieving higher specificity and less adverse effects. This concept utilizes the adoptive transfer of autologous T cells that have been retrovirally transduced ex vivo to express and deliver immunoregulatory gene products to sites of autoimmune inflammation. In the animal model of collagen-induced autoimmune polychondritis ear disease (CIAPED), the adoptive transfer of IL-12p40-expressing collagen type II (CII)-specific CD4+ T-cell hybridomas resulted in a significantly lower disease incidence and severity compared with untreated or vector-only-treated animals...
May 3, 2017: ORL; Journal for Oto-rhino-laryngology and its related Specialties
https://www.readbyqxmd.com/read/28462722/the-regulatory-functions-of-a-new-tetrapeptide-from-the-bursa-of-fabricius-on-aiv-vaccine-immunization-and-antibody-production
#11
Man Man Zong, Chuan Jie Zhou, Yuan Nan Yu, Yang Zheng, Xiu Li Feng, Pu Yan Chen, Mei Yang
The bursa of Fabricius (BF) is the acknowledged humoral central immune organ and plays vital role on B lymphocyte development. In the study, a new tetrapeptide of GERA isolated from BF was named bursal peptide tetrapeptide (BP4). To investigate the immunomodulatory function of BP4 on antigen-specific immune responses, BALB/c mice were immunized with inactivated influence virus (AIV, H9N2 subtype) vaccine and BP4 at three experimental dosages. The results showed that BP4 enhanced the increased AIV-specific antibody and cytokine production, and modified T cell subtype population...
April 25, 2017: Protein and Peptide Letters
https://www.readbyqxmd.com/read/28461659/direct-and-indirect-cell-based-enzyme-linked-immunosorbent-assay
#12
Thomas O Kohl, Carl A Ascoli
The direct cell-based enzyme-linked immunosorbent assay (ELISA) has developed into a popular alternative immunoassay for the rapid detection of expressed cell-surface antigens or receptors. It is used to determine cell surface antigen expression profiles using existing reporter-labeled antibodies. The target specificity of newly developed antibodies can be determined by the indirect approach, which screens hybridoma supernatants for antibody reactivity against the target antigen of interest. The cell-based ELISA acts as a surrogate for the identification of immunohistochemistry-reactive (IHC) antibodies and aids in hybridoma-derived antibody identification, which is used for further ELISA development...
May 1, 2017: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/28458052/production-of-monoclonal-antibodies-and-development-of-a-quantitative-immuno-polymerase-chain-reaction-assay-to-detect-and-quantify-recombinant-glutathione-s-transferase
#13
Abud Je, Luque Eh, Ramos Jg, Rodriguez Ha
GST-tagged proteins are important tools for the production of recombinant proteins. Removal of GST tag from its fusion protein, frequently by harsh chemical treatments or proteolytic methods, is often required. Thus, the monitoring of the proteins in tag-free form requires a significant effort to determine the remnants of GST during purification process. In the present study, we developed both a conventional enzyme-linked immunosorbent assay (ELISA) and an immuno-polymerase chain reaction (IPCR) assay, both specific for detection of recombinant GST (rGST)...
April 27, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28424886/detection-of-koi-herpesvirus-khv-using-a-monoclonal-antibody-against-cyprinus-carpio-igm
#14
Yingying Li, Shucheng Zheng, Qing Wang, Sven M Bergmann, Weiwei Zeng, Yingying Wang, Chun Liu, Cunbin Shi
Koi herpesvirus disease (KHVD) is associated with high mortality in both common carp and koi carp (Cyprinus carpio L.) worldwide. The indirect detection of fish viruses based on the identification of antibodies has emerged as a practical and reliable means of diagnosis. Thus, it is important to create monoclonal antibodies (MAbs) against carp IgM. By using hybridoma-monoclonal antibody technology, one hybridoma cell line secreting MAbs against IgM from carp was established. In western blot analysis, the secreted MAb from cell line A5-E10 recognized the heavy chain of IgM from common carp or koi but did not react with immunoglobulins from three different fish species: grass carp (Ctenopharyngodon idella), tilapia (Oreochromis mossambicus) and Mandarin fish (Siniperca chuatsi)...
April 19, 2017: Archives of Virology
https://www.readbyqxmd.com/read/28421663/cloning-and-plant-based-production-of-antibody-mc10e7-for-a-lateral-flow-immunoassay-to-detect-4-arginine-microcystin-in-fresh-water
#15
Stanislav Melnik, A-C Neumann, R Karongo, S Dirndorfer, Martin Stübler, Verena Ibl, R Niessner, Dietmar Knopp, Eva Stoger
Antibody MC10E7 is one of a small number of monoclonal antibodies that bind specifically to [Arg4]-microcystins and it can be used to survey natural water sources and food samples for algal toxin contamination. However, the development of sensitive immunoassays in different test formats, particularly user-friendly tests for on-site analysis, requires a sensitive but also cost-effective antibody. The original version of MC10E7 was derived from a murine hybridoma, but we determined the sequence of the variable regions using the peptide mass-assisted cloning strategy and expressed a scFv (single-chain variable fragment) format of this antibody in yeast and a chimeric full size version in leaves of Nicotiana tabacum and N...
April 19, 2017: Plant Biotechnology Journal
https://www.readbyqxmd.com/read/28417626/production-and-characterization-of-monoclonal-antibody-against-recombinant-virus-coat-protein-cp42
#16
Naeimeh Shibaei, Jafar Majidi, Khadijeh Razavi, Ali Asghar Karkhane, Nemat Sokhandan-Bashir, Leili Aghebati-Maleki
There are many studies related to the production of a ELISA kit for diagnosing virus infections. However, production of most kits depends on purification of whole virus particles, which involves the use of costly equipment and reagents. The purpose of this study was to check out if the anti-CP42 antibodies could be used as a diagnostic assay for detection of Grapevine fanleaf Virus (GFLV). In this study, recombinant GFLV coat protein gene related to selected antigenic determinants was inserted into pET-28a bacterial expression vector and the construct (pET-28a CP42) was cloned into E...
February 2017: Iranian Journal of Allergy, Asthma, and Immunology
https://www.readbyqxmd.com/read/28414197/engineering-of-a-self-adjuvanted-itep-delivered-ctl-vaccine
#17
Shuyun Dong, Tiefeng Xu, Peng Wang, Peng Zhao, Mingnan Chen
Cytotoxic T lymphocyte (CTL) epitope peptide-based vaccines are widely used in cancer and infectious disease therapy. We previously generated an immune-tolerant elastin-like polypeptides (iTEPs)-based carrier to deliver a peptide CTL vaccine and enhance the efficiency of the vaccine. To further optimize the vaccine carrier, we intended to potentiate its function by designing an iTEP-based carrier that was able to deliver adjuvant and a vaccine epitope as one molecule. Thus, we fused a 9-mer H100, a peptide derived from the high-mobility group box 1 protein (HMGB1) that could induce activation of dendritic cells (DCs), with an iTEP polymer to generate a new iTEP polymer named H100-iTEP...
April 17, 2017: Acta Pharmacologica Sinica
https://www.readbyqxmd.com/read/28414151/targeted-antigen-delivery-to-dendritic-cell-via-functionalized-alginate-nanoparticles-for-cancer-immunotherapy
#18
Chuangnian Zhang, Gaona Shi, Ju Zhang, Huijuan Song, Jinfeng Niu, Shengbin Shi, Pingsheng Huang, Yanming Wang, Weiwei Wang, Chen Li, Deling Kong
The purpose of the present study was to identify an "easy-to-adopt" strategy to enhance immune responses using functionalized alginate (ALG) nanoparticles (MAN-ALG/ALG=OVA NPs), which were prepared by CaCl2 cross-linking of two different types of ALG. The mannose (MAN) modified ALG (MAN-ALG) was used for dendritic cell targeting. The other component, composed of ovalbumin (OVA), a model antigen, is conjugated to ALG (ALG=OVA) via pH sensitive Schiff base bond. Grafting of alginate was demonstrated by FT-IR and (1)H NMR, while the morphological structure, particle size, Zeta potential of MAN-ALG/ALG=OVA NPs were measured using TEM and DLS...
April 13, 2017: Journal of Controlled Release: Official Journal of the Controlled Release Society
https://www.readbyqxmd.com/read/28414024/novel-high-throughput-cell-based-hybridoma-screening-methodology-using-the-celigo-image-cytometer
#19
Haohai Zhang, Leo Li-Ying Chan, William Rice, Nasim Kassam, Maria Serena Longhi, Haitao Zhao, Simon C Robson, Wenda Gao, Yan Wu
Hybridoma screening is a critical step for antibody discovery, which necessitates prompt identification of potential clones from hundreds to thousands of hybridoma cultures against the desired immunogen. Technical issues associated with ELISA- and flow cytometry-based screening limit accuracy and diminish high-throughput capability, increasing time and cost. Conventional ELISA screening with coated antigen is also impractical for difficult-to-express hydrophobic membrane antigens or multi-chain protein complexes...
April 14, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28412548/internal-image-anti-idiotypic-antibody-a-new-strategy-for-the-development-a-new-category-of-prolactin-receptor-prlr-antagonist
#20
Hainan Lan, Pan Hong, Ruonan Li, Suo L, Shan Anshan, Steven Li, Xin Zheng
Over the past decades, a number of prolactin receptor (PRLR) antagonists have been developed, which can be divided into two categories, PRLR analogue and anti-PRLR antibody. However, until now, there have been no commercially available PRLR antagonists. Here, we described a new approach for the preparation of PRLR antagonist, namely internal image anti-idiotypic antibody strategy. The hybridoma technique was used to generate anti-idiotypic antibodies to PRL. Competitive ELISA, competitive receptor-binding analysis and immunofluorescence assay (IFA) were then used to screen and characterize anti-idiotypic antibodies to PRL...
April 13, 2017: Molecular Immunology
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