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Bian C.,Chinese Academy of Sciences | Zhang F.,Shanghai JiaoTong University | Wang F.,Shanghai JiaoTong University | Ling Z.,CAS Institut Pasteur of Shanghai | And 16 more authors.
Acta Biochimica et Biophysica Sinica | Year: 2010

DNA immunization is an efficient method for high-affinity monoclonal antibody generation. Here, we describe the generation of several high-quality monoclonal antibodies (mAbs) against retinol-binding protein 4 (RBP4), an important marker for kidney abnormality and dysfunction, with a combination method of DNA priming and protein boost. The mAbs generated could bind to RBP4 with high sensitivity and using these mAbs, an immunocolloidal gold fast test strip was constructed. The strip can give a result in <5 min and is very sensitive with a detection limit of about 1 ng/ml. A small-scale clinical test revealed that the result of this strip was well in accordance with that of an enzyme-labeled immunosorbent assay kit currently available on the market. Consequently, it could be useful for more convenient and faster RBP4 determination in the clinic. © 2010 The Author.


Zhuang X.,Chinese Academy of Sciences | Sun Y.,Chinese Academy of Sciences | Ling Z.,CAS Institut Pasteur of Shanghai | Dong Q.,Chinese Academy of Sciences | And 12 more authors.
Comptes Rendus Chimie | Year: 2011

Monoclonal antibodies are needed for the development of effective, accurate and sensitive immunoassays against the H5N1 avian influenza. Here we report the generation of monoclonal antibodies targeting Escherichia coli expressed hemagglutinin of HA1 domain from H5N1 influenza virus. The monoclonal antibodies can be used to establish a sandwich ELISA assay that is able to detect HA from H5 subtype virus lysate but not from H9 subtype. A subtype-conserved new epitope was also identified which is located between the amino acid residues N80 to A99. This work would be helpful for the development of a diagnosis kit for the H5N1 avian influenza. © 2011 Académie des sciences. Published by Elsevier Masson SAS. All rights reserved.


Sun Y.,CAS Shanghai Institutes for Biological Sciences | Bian C.,CAS Shanghai Institutes for Biological Sciences | Xu K.,CAS Institut Pasteur of Shanghai | Hu W.,CAS Institut Pasteur of Shanghai | And 19 more authors.
PLoS ONE | Year: 2010

Background: The 2009 swine-origin influenza virus (S-OIV) H1N1 pandemic has caused more than 18,000 deaths worldwide. Vaccines against the 2009 A/H1N1 influenza virus are useful for preventing infection and controlling the pandemic. The kinetics of the immune response following vaccination with the 2009 A/H1N1 influenza vaccine need further investigation. Methodology/Principal Findings: 58 volunteers were vaccinated with a 2009 A/H1N1 pandemic influenza monovalent split-virus vaccine (15 mg, single-dose). The sera were collected before Day 0 (pre-vaccination) and on Days 3, 5, 10, 14, 21, 30, 45 and 60 post vaccination. Specific antibody responses induced by the vaccination were analyzed using hemagglutination inhibition (HI) assay and enzyme-linked immunosorbent assay (ELISA). After administration of the 2009 A/H1N1 influenza vaccine, specific and protective antibody response with a major subtype of IgG was sufficiently developed as early as Day 10 (seroprotection rate: 93%). This specific antibody response could maintain for at least 60 days without significant reduction. Antibody response induced by the 2009 A/H1N1 influenza vaccine could not render protection against seasonal H1N1 influenza (seroconversion rate: 3% on Day 21). However, volunteers with higher pre-existing seasonal influenza antibody levels (pre-vaccination HI titer ≥1:40, Group 1) more easily developed a strong antibody protection effect against the 2009 A/H1N1 influenza vaccine as compared with those showing lower pre-existing seasonal influenza antibody levels (pre-vaccination HI titer, <:40, Group 2). The titer of the specific antibody against the 2009 A/H1N1 influenza was much higher in Group 1 (geometric mean titer: 146 on Day 21) than that in Group 2 (geometric mean titer: 70 on Day 21). Conclusions/Significance: Recipients could gain sufficient protection as early as 10 days after vaccine administration. The protection could last at least 60 days. Individuals with a stronger pre-existing seasonal influenza antibody response may have a relatively higher potential for developing a stronger humoral immune response after vaccination with the 2009 A/ H1N1 pandemic influenza vaccine. © 2010 Sun et al.


Kucukerden M.,Istanbul University | Kucukerden M.,University of Texas Medical Branch | Huda R.,University of Texas Medical Branch | Tuzun E.,Istanbul University | And 14 more authors.
Journal of Neuroimmunology | Year: 2016

Sera of myasthenia gravis (MG) patients with muscle-specific receptor kinase-antibody (MuSK-Ab) predominantly display the non-complement fixing IgG4 isotype. Similarly, mouse IgG1, which is the analog of human IgG4, is the predominant isotype in mice with experimental autoimmune myasthenia gravis (EAMG) induced by MuSK immunization. The present study was performed to determine whether IgG1 anti-MuSK antibody is required for immunized mice to develop EAMG. Results demonstrated a significant correlation between clinical severity of EAMG and levels of MuSK-binding IgG1 +, IgG2 + and IgG3 + peripheral blood B cells in MuSK-immunized wild-type (WT) mice. Moreover, MuSK-immunized IgG1 knockout (KO) and WT mice showed similar EAMG severity, serum MuSK-Ab levels, muscle acetylcholine receptor concentrations, neuromuscular junction immunoglobulin and complement deposit ratios. IgG1 and IgG3 were the predominant anti-MuSK isotypes in WT and IgG1 KO mice, respectively. These observations demonstrate that non-IgG1 isotypes can mediate MuSK-EAMG pathogenesis. © 2016 Elsevier B.V.


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