Differentiation among the Vibrio cholerae serotypes O1, O139, O141 and non-O1, non-O139, non-O141 using specific monoclonal antibodies with dot blotting

Pengsuk, Chalinan; Longyant, Siwaporn; Rukpratanporn, Sombat; Chaivisuthangkura, Parin; Sridulyakul, Pattarin; Sithigorngul, Paisarn

doi:10.1016/j.mimet.2011.07.022

Cited by 16 publications

(16 citation statements)

References 41 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…generally used to measure and identify the contents of antigens in multivalent vaccines. To date, monoclonal antibodies (mAbs) directed against V. cholerae O1 have been obtained by immunization of mice with the LPS or strain(s) of Ogawa and Inaba serotypes using hybridoma technology [8,[10][11][12][13][14][15][16][17][18][19][20][21]. Most of the mAbs reacted with both Inaba and Ogawa serotypes, due to the minor structural differences in these serotypes, and few of them were Ogawa-or Inaba-serotype specific [8,14].…”

Section: Vibrio Cholerae Cause Of the Life-threatening Diarrheal Dismentioning

confidence: 99%

Generation and Characterization of Monoclonal Antibodies to the Ogawa Lipopolysaccharide of Vibrio cholerae O1 from Phage-Displayed Human Synthetic Fab Library

Kim

Hong

Choi

et al. 2020

J. Microbiol. Biotechnol.

View full text Add to dashboard Cite

Vibrio cholerae , cause of the life-threatening diarrheal disease cholera, can be divided into different serogroups based on the structure of its lipopolysaccharide (LPS), which consists of lipid-A, corepolysaccharide and O-antigen polysaccharide (O-PS). The O1 serogroup, the predominant cause of cholera, includes two major serotypes, Inaba and Ogawa. These serotypes are differentiated by the presence of a single 2- O -methyl group in the upstream terminal perosamine of the Ogawa O-PS, which is absent in the Inaba O-PS. To ensure the consistent quality and efficacy of the current cholera vaccines, accurate measurement and characterization of each of these two serotypes is highly important. In this study, we efficiently screened a phage-displayed human synthetic Fab library by bio-panning against Ogawa LPS and finally selected three unique mAbs (D9, E11, and F7) that specifically react with Ogawa LPS. The mAbs bound to Vibrio cholerae vaccine in a dose-dependent fashion. Sequence and structure analyses of antibody paratopes suggest that IgG D9 might have the same fine specificity as that of the murine mAbs, which were shown to bind to the upstream terminal perosamine of Ogawa O-PS, whereas IgGs F7 and E11 showed some different characteristics in the paratopes. To our knowledge, this study is the first to demonstrate the generation of Ogawa-specific mAbs using phage display technology. The mAbs will be useful for identification and quantification of Ogawa LPS in multivalent V. cholerae vaccines.

show abstract

Section: Vibrio Cholerae Cause Of the Life-threatening Diarrheal Dismentioning

confidence: 99%

Generation and Characterization of Monoclonal Antibodies to the Ogawa Lipopolysaccharide of Vibrio cholerae O1 from Phage-Displayed Human Synthetic Fab Library

Kim

Hong

Choi

et al. 2020

J. Microbiol. Biotechnol.

View full text Add to dashboard Cite

show abstract

“…The interesting point is that, they screened their positive cultures by dot blotting and western immunoassay against each of the 5 isolates of V. cholerae that were used for the immunization of mice before cell fusion (Pengsuk et al, 2011).…”

Section: Ajimentioning

confidence: 99%

“…In this regard, it seems that immunoassay could be the simplest and easy way with enough specificity and sensitivity in recognizing different species of V. cholerae. Pensuk and his colleagues have demonstrated a rapid method for diagnosis of various V. cholerae based on specific monoclonal antibodies (Pengsuk et al, 2010;2011;Prompamorn et al, 2013). The aim of this study was to developed mouse monoclonal and rabbit polyclonal antibodies against recombinant OMPw protein that might be useful for immunodiagnostic of V. cholerae.…”

Section: Introductionmentioning

confidence: 99%

Production and Characterization of Monoclonal and Polyclonal Antibody Against Recombinant Outer Membrane Protein

Fasihi-Ramandi¹

2014

American Journal of Immunology

View full text Add to dashboard Cite

There are many studies related to immunological and molecular methods for diagnosis of Vibrio cholera (V. cholerae). However, most assays dependent on enrichment of culture of bacteria, which need more time and involves the use of costly equipment and reagents. In this study Balb/c mice were immunized with recombinant Outer Membrane Protein (rOMPw) of vibrio cholerae and splenocytes of hyper immunized mice were fused with murine myeloma Sp2/0 cells. Positive hybridomas were selected by ELISA using rOMPw as coating antigen. The monoclonal antibodies from ascitic fluids were purified and its reaction with rOMPw was assessed by ELISA. Polyclonal antibodies were also produced by immunization of rabbits with the above mentioned antigen. The rabbit sera was affinity purified using Hi-Trap protein G column. The result showed that monoclonal antibody specific to rOMPw has been successfully generated. The monoclonal antibody reacted with recombinant OMPw in ELISA and immunonoblat method. Rabbit polyclonal antibody was also bound to rOMPw by ELISA. The results of agglutination test with whole bacteria also showed that both mouse monoclonal and rabbit polyclonal antibodies reacted with whole vibrio cholera but not other related bacteria. The purpose of this study was to check out if anti OMPw antibodies could use as diagnostic assay for detection of V. cholerae. Our results demonstrated that anti recombinant OMPw monoclonal and polyclonal antibodies are able to diagnose whole bacteria in pure culture using agglutination test but not by home made immunochromatic strip test.

show abstract

“…LFIC is a simple and widely used technique for the detection of the presence (or absence) of a target analyte in sample mixtures without the need for extensive sample preparation steps [ 17 – 25 ]. A lateral-flow apparatus consisting of a simple membrane with a capture zone and pads through which reagents can be introduced allows the reaction to be completed in a relatively short time (~ 15 minutes) and further enables the in-situ separation of unreacted components via one-step analysis, a key advantage of this format.…”

Section: Introductionmentioning

confidence: 99%

Point-of-care test for cervical cancer in LMICs

et al. 2016

View full text Add to dashboard Cite

Cervical cancer screening using Papanicolaou's smear test has been highly effective in reducing death from this disease. However, this test is unaffordable in low- and middle-income countries, and its complexity has limited wide-scale uptake. Alternative tests, such as visual inspection with acetic acid or Lugol's iodine and human papillomavirus DNA, are sub-optimal in terms of specificity and sensitivity, thus sensitive and affordable tests with high specificity for on-site reporting are needed. Using proteomics and bioinformatics, we have identified valosin-containing protein (VCP) as differentially expressed between normal specimens and those with cervical intra-epithelial neoplasia grade 2/3 (CIN2/CIN3+) or worse. VCP-specific immunohistochemical staining (validated by a point-of-care technology) provided sensitive (93%) and specific (88%) identification of CIN2/CIN3+ and may serve as a critical biomarker for cervical-cancer screening. Future efforts will focus on further refinements to enhance analytic sensitivity and specificity of our proposed test, as well as on prototype development.

show abstract

Differentiation among the Vibrio cholerae serotypes O1, O139, O141 and non-O1, non-O139, non-O141 using specific monoclonal antibodies with dot blotting

Cited by 16 publications

References 41 publications

Generation and Characterization of Monoclonal Antibodies to the Ogawa Lipopolysaccharide of Vibrio cholerae O1 from Phage-Displayed Human Synthetic Fab Library

Generation and Characterization of Monoclonal Antibodies to the Ogawa Lipopolysaccharide of Vibrio cholerae O1 from Phage-Displayed Human Synthetic Fab Library

Production and Characterization of Monoclonal and Polyclonal Antibody Against Recombinant Outer Membrane Protein

Point-of-care test for cervical cancer in LMICs

Contact Info

Product

Resources

About