Use of monoclonal antibodies for detection of infectious pancreatic necrosis virus by the enzyme-linked immunosorbent assay (ELISA)

Domínguez, José Luis; Hedrick, Ronald P.; Sánchez-Vizcaíno, J M

doi:10.3354/dao008157

Cited by 35 publications

(11 citation statements)

References 12 publications

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“…For labeling, mAbs were purified from ascitic fluid by affinity chromatography on Protein ASepharose CL 4B (Pharmacia, Sweeden). Biotinylation of mAbs was performed as previously described [4].…”

Section: Monoclonal Antibody Productionmentioning

confidence: 99%

Characterization of a novel activation antigen on porcine lymphocytes recognized by monoclonal antibody 5A6/8

Doménech

Bullido²,

Álvarez³

et al. 2004

Vet. Res.

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-In this report, we describe the characterization of a novel activation antigen on porcine lymphocytes recognized by mAb 5A6/8. This antigen was detected on B and T cells 24 h after treatment with various stimuli. It was also found on alveolar macrophages, and at low levels on untreated monocytes. MAb 5A6/8 precipitated two bands of 45 and 50 kDa under non-reducing conditions, and of 22 and 28 kDa under reducing conditions. The cellular distribution, expression kinetics and/or molecular size of the 5A6/8 antigen differ from those of other known lymphocyte activation antigens. MAb 5A6/8 was able to inhibit lymphocyte proliferative responses driven by different stimuli, suggesting a role for this molecule in the events that lead to lymphocyte activation. activation antigen / lymphocyte / monoclonal antibody / monocyte / pig

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Section: Monoclonal Antibody Productionmentioning

confidence: 99%

Characterization of a novel activation antigen on porcine lymphocytes recognized by monoclonal antibody 5A6/8

Doménech

Bullido²,

Álvarez³

et al. 2004

Vet. Res.

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“…Antigen-affinity-purified Abs are free of the problen~s mentioned above but they might be difficult to obtain because of the large amounts of purified virus needed to make the affinity columns. MAbs would probably be the best alternative since they have already been obtained and characterized against IPNV (CaswellReno et al 1986, Wolski et al 1986, Dominguez et al 1990), VHSV (Lorenzen et al 1988, Mourton et al 1990 and IHNV (Schultz et al 1985, Winton et al 1988.…”

Section: Amplification Of Viral Content Of Samplesmentioning

confidence: 99%

“…By neutralization it was also possible to differentiate 3 VHSV serotypes (Le Berre et al 1977), but no cross-neutralization between IHNV and VHSV has yet been demonstrated (McAllister et al 1974), nor have different IHNV serotypes been described (Engelking et al 1991). Neutralizing MAbs have been difficult to obtain for IPNV (Dominguez et al 1990), VHSV (Lorenzen et al 1990, Sanz et al unpubl. ) and IHNV (Winton et al 1988.…”

Section: Techniques For Identification Of Viral Proteins Neutralizationmentioning

confidence: 99%

“…Moribund fry from a natural infection with 90 % mortality showed a high value by ELISA. A sensitivity increase was claimed by Hattori et al (1984) to allow detection of 104 TCIDso ml-l and by Rodak et al (1988) to allow detection of lo3 TCIDSo ml-l. Dominguez et al (1990) described the use of MAbs with similar detection levels corresponding to 10 ng of purified virus ml-'. Rather than on new methodological approaches, however, these differences seem to depend on the quality of the Abs, the amount of Ab bound to the solid phase or the presence of variable concentrations of non-infective viral particles in the samples tested (Table 2).…”

Section: Agglutinationmentioning

confidence: 99%

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Techniques for diagnosing viral diseases of salmonid fish

Sanz¹,

Coll²

1992

Dis. Aquat. Org.

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Cell culture for amplification and the techniques most used for identification of salmonid viruses -neutralization, immunofluorescence and, to a lesser extent, immunoperoxldase, complen~ent fixation, agglutination, electron microscopy, immunodifussion or radioimmunoassay -may soon b e replaced by other techniques such as enzyme immunoassays (immunodot and enzyme-linked immunosorbent assay, ELISA) and hybridization with DNA probes. It is expected that developments in monoclonal antibodies (MAbs) and amplification by the polymerase chain reaction (PCR) will increase sensitivity of enzyme immunoassays and DNA hybridizations, respectively. Some of these new methods should provide detection of the low levels of virus present in adult carriers and perhaps in eggs (although this is more complicated). Other &agnostic methods, such as measurement of virus-specific salmonid immunoglobulins (Igs) by ELISA or stimulation of immunological cellular memory by in vitro CO-culture of salmonid lymphocytes with viral proteins, could also b e further developed.

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“…For labeling, mAbs were purif ied from hybridoma cell culture supernatants by affinity chromatography with Protein G-Sepharose CL 4B (Pharmacia, Sweden). Biotinylation of mAbs was performed as described previously (Domínguez et al, 1990).…”

Section: Antibodiesmentioning

confidence: 99%

Cloning and expression of porcine CD163: its use for characterization of monoclonal antibodies to porcine CD163 and development of an ELISA to measure soluble CD163 in biological fluids

Pérez¹,

Ezquerra²,

Ortuño³

et al. 2008

Span. j. agric. res.

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CD163 is a member of the family of proteins with scavenger receptor cysteine-rich domains, whose expression is restricted to monocytes and macrophages. It functions as a receptor for haemoglobin/haptoglobin complexes and it has also been implicated in the regulation of inflammatory processes. Treatment of monocytes/macrophages with phorbol esters, LPS or cross-linking of Fc gamma receptors, causes shedding of CD163 from cell surface by a protease-dependent mechanism. The level of soluble CD163 (sCD163) in serum and other body fluids has been considered a useful marker of macrophage activation. In addition, porcine CD163 has been shown to play a role in the infection of monocytes/macrophages by the African swine fever virus. The porcine CD163 cDNA has been cloned, and expressed in transfected CHO cells. Clones of CHO cells stably expressing porcine CD163 have been used for the characterization of three new mAbs against porcine CD163. Using two of these mAbs, that bind to different epitopes on CD163 molecule, a sandwich enzyme-linked immunoassay (ELISA) has been developed to measure levels of sCD163 in porcine sera and biological fluids. The assay was calibrated using lysates of CD163 transfectants, showing a sensitivity of 10 5 cells mL -1 , that allowed to detect sCD163 in sera and in culture supernatants of activated alveolar macrophages. This assay can be a useful method of monitoring the degree of activation of macrophages in a variety of inflammatory and infectious diseases of swine.Additional key words: macrophage, SRCR family, swine. ResumenClonación y expresión de CD163 porcino: aplicación en la caracterización de anticuerpos monoclonales frente a CD163 porcino, y desarrollo de un ELISA para cuantificar CD163 soluble en fluidos biológicos CD163 es un miembro de la familia de proteínas con dominios de receptores «scavenger» ricos en cisteína, con una expresión restringida a células del linaje monocítico. Se ha demostrado su función como receptor de complejos hemoglobina/haptoglobina, y se le ha implicado en la regulación de procesos inflamatorios. El tratamiento de monocitos/macrófagos con ésteres de forbol, LPS, o mediante entrecruzamiento de receptores Fc gamma, provoca la liberación de CD163 de la superficie celular por un mecanismo dependiente de proteasas. La cantidad de CD163 soluble (sCD163) en suero y otros fluidos corporales se considera un indicador de la activación de los macrófagos. Recientemente, nuestro grupo demostró que CD163 está implicado en la infección de monocitos/macrófagos porcinos por el virus de la peste porcina africana. En este trabajo se describe la clonación del cDNA de CD163 porcino, y su expresión en células CHO transfectadas. Se obtuvieron clones de células CHO transfectadas que expresan de forma estable la molécula CD163 porcina, y se utilizaron para caracterizar tres nuevos anticuerpos monoclonales que reconocen específicamente CD163 porcino. El uso de dos de estos anticuerpos, que reconocen epítopos diferentes en CD163 porcino, ha permitido desarrollar un ensayo i...

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Use of monoclonal antibodies for detection of infectious pancreatic necrosis virus by the enzyme-linked immunosorbent assay (ELISA)

Cited by 35 publications

References 12 publications

Characterization of a novel activation antigen on porcine lymphocytes recognized by monoclonal antibody 5A6/8

Characterization of a novel activation antigen on porcine lymphocytes recognized by monoclonal antibody 5A6/8

Techniques for diagnosing viral diseases of salmonid fish

Cloning and expression of porcine CD163: its use for characterization of monoclonal antibodies to porcine CD163 and development of an ELISA to measure soluble CD163 in biological fluids

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