Production, characterisation and applicability of monoclonal antibodies to European eel (Anguilla anguilla L., 1758) immunoglobulin

Heijden, M.H.T. van der; Rooijakkers, J.B.M.A.; Booms, G.H.R.; Rombout, J.H.W.M.; Boon, J.H.

doi:10.1016/0165-2427(94)05335-p

Cited by 47 publications

(33 citation statements)

References 17 publications

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“…Immunoblot analysis of normal eel blood incubated with positive eel sera revealed only a weak band of about 67 kDa, which probably represents the heavy chain of eel immunoglobulin (Van der Heijden et al 1995, Hung et al 1996. This band was not detectable in the antigen preparation of the parasite intestinal content, indicating that the eel immunoglobulin in the adult parasite gut was probably enzymatically decomposed.…”

Section: Discussionmentioning

confidence: 99%

“…As secondary and tertiary antibodies, monoclonal mouse IgG specific for eel immunoglobulin heavy chain (WEI 1, Van der Heijden et al 1995) diluted 1:500 in PBS-T-BSA and sheep anti-mouse IgG conjugated with horseradish peroxidase (AP271, The Binding Site, England) diluted 1:1000 in PBS-T-BSA were used, respectively. Both the secondary and tertiary antibody were incubated for 45 min at 37°C.…”

Section: Source Of Antigens Adult Anguillicola Crassus Andmentioning

confidence: 99%

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Evaluation of an ELISA and immunoblotting for studying the humoral immune response in Anguillicola crassus infected European eel Anguilla anguilla

Knopf¹,

Naser²,

Heijden³

et al. 2000

Dis. Aquat. Org.

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The applicability of an enzyme-linked immunosorbent assay (ELISA) for the detection of anguillicolosis in feral eels was examined using a crude antigen preparation from the body wall of adult Anguillicola crassus. The screening consisted of samples from 100 feral European eels Anguilla anguilla. As a reference the actual status of infection was determined by dissection of the eels' swimbladders. The ELISA results were compared with a background value calculated from the results obtained from 43 non-infected farm eels. The screened samples had a high prevalence of A. crassus (83%); however, the specificity and the negative predictive value of the ELISA were low compared to the high positive predictive value. Nonetheless, the reproducibility (precision) of the test was satisfactory, and for the non-infected reference group specificity was 97.7%. Although the ELISA, as used in the present study, is not applicable for diagnostic purposes, it represents a useful tool for the investigation of the specific humoral immune response of eels against A. crassus under controlled experimental conditions. Immunoblots using crude antigen preparations from different parts of adult A. crassus as well as a crude somatic third-stage (L 3 ) antigen preparation illustrated that only antigens associated with the body wall of adult A. crassus are potentially suitable for diagnostic purposes. Despite the fact that antibodies against Raphidascaris acus cross-reacted with 3 body wall antigens of A. crassus, the most encouraging results were obtained with the antigen preparation from the outer cuticle of adult A. crassus which yielded a conspicuous, broad band at about 100 kDa.

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Section: Discussionmentioning

confidence: 99%

Section: Source Of Antigens Adult Anguillicola Crassus Andmentioning

confidence: 99%

Evaluation of an ELISA and immunoblotting for studying the humoral immune response in Anguillicola crassus infected European eel Anguilla anguilla

Knopf¹,

Naser²,

Heijden³

et al. 2000

Dis. Aquat. Org.

View full text Add to dashboard Cite

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“…During the last decade, MAbs have been produced to Ig of a number of teleost species (DeLuca et al, 1983;Secombes et al, 1983;Thuvander et al, 1990;Navarro et al, 1993;Sanchez et al, 1993). Most of these antibodies recognize the heavy (H) chain of Ig, whereas a few were obtained against the light (L) chain (Lobb et al, 1984;Sanchez and Dominguez, 1991;van der Heijden et al, 1995). This has greatly contributed to an improved understanding of the architecture and functioning of the immune system of teleost fish in general and the corresponding species in particular.…”

Section: Introductionmentioning

confidence: 99%

Characterization of Monoclonal Antibodies against Heavy and Light Chains of Flounder (Paralichthys olivaceus) Immunoglobulin

Jang

Woo²,

Cho³

et al. 2004

BMB Reports

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“…Eel sera diluted 1:100 in ELISA buffer were preheated for 30 min at 45°C and were incubated on the virus coating. Antibodies bound to HVA were detected by successive incubation with a monoclonal antibody specific for the heavy chain of eel immunoglobulin (a 1:30 dilution of cell culture medium of hybridoma WEI-1 in ELISA buffer; van der Heijden et al 1995) and peroxidase conjugated antibody specific for mouse immunoglobulin (a 1:500 dilution of rabbit anti-mouse serum conjugated to peroxidase; Dakopatts, Denmark, Cat. number P161).…”

Section: Methodsmentioning

confidence: 99%

Persistence of herpesvirus of eel Herpesvirus anguillae in farmed European eel Anguilla anguilla

Nieuwstadt¹,

Dijkstra²,

Haenen³

2001

Dis. Aquat. Org.

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Herpesvirus of eel Herpesvirus anguillae (HVA) was isolated repeatedly from farmed eel of an outwardly healthy stock, but virus isolation was much greater in an experimental group of fish that were injected with dexamethasone. The results suggest that HVA can establish a latent infection in eel. Previous exposure of these eels to HVA virus was shown by detection of HVA-specific antibodies. These eels did not show clinical signs after a secondary infection with HVA. Tracing of seropositive eel stocks, which had previous contact with HVA, and of HVA carrier fish can be useful to control disease outbreaks due to HVA infection. KEY WORDS: Herpesvirus · Eel · Latency · Dexamethasone Resale or republication not permitted without written consent of the publisherDis Aquat Org 45: [103][104][105][106][107] 2001 MATERIALS AND METHODS Cells, virus, antiserum. EK-1 cells (Chen et al. 1982) were used in the isolation and propagation of HVA throughout this study. Cells were routinely cultured in sterile plastic flasks in Leibovitz-L15 medium (Life Technologies BV, Breda, The Netherlands) supplemented with 5% foetal bovine serum, antibiotics and 0.075 to 0.15% (w/v) bicarbonate. Cultures were incubated at 26°C in a CO 2 incubator. The Dutch isolate 500138 of HVA was propagated in EK-1 cells, and virus was concentrated and purified from cell culture medium by centrifugation for 4 h at 53 000 × g through 25% (w/w) sucrose in TEN buffer (10 mM Tris, 150 mM NaCl, 1 mM EDTA; pH 7.4). The virus pellet was resuspended in TE buffer (10 mM Tris. HCl, 1 mM EDTA, pH 7.4) and virus protein concentration was determined using the BCA (bicinchoninic acid) protein assay with bovine serum albumin as a standard (Pierce, Rockford, IL, USA). To obtain specific antiserum, rabbits were immunised 3 times both intramuscularly and subcutaneously with a dose of 1 mg virus protein in complete or incomplete Freund adjuvans. The rabbit immune serum had a titre of 1:1060 in a virus neutralisation test using 150 TCID 50 of HVA. The rabbit anti-HVA serum was used to type virus isolates, which showed a cytopathic effect characteristic of HVA.Fish. Eighty outwardly healthy eels ranging in weight from 150 to 200 g were purchased from an eel farm. Fishes were kept at 23°C in six 200 l glass aquariums half filled with tap water, which was aerated continuously and changed for fresh water each day. Water temperature was recorded each day. Eels were offered a hiding place in these aquariums and were subjected to a 12/12 h light/dark cycle. They were fed at a maintenance level with commercial dry pellet feed. For individual identification of fishes from which blood samples were collected weekly, 26 fishes in 2 aquariums were tagged by scratching their skin.Experimental design. Eels were placed in 6 aquariums (A to F) in groups of 13 or 14 eels. First blood samples were collected from the caudal vein from all fishes. The experiment was started on Day 0, which was 4 d after eels arrived at our institute. Fishes in Aquariums A, B, and C were untreated contr...

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Production, characterisation and applicability of monoclonal antibodies to European eel (Anguilla anguilla L., 1758) immunoglobulin

Cited by 47 publications

References 17 publications

Evaluation of an ELISA and immunoblotting for studying the humoral immune response in Anguillicola crassus infected European eel Anguilla anguilla

Evaluation of an ELISA and immunoblotting for studying the humoral immune response in Anguillicola crassus infected European eel Anguilla anguilla

Characterization of Monoclonal Antibodies against Heavy and Light Chains of Flounder (Paralichthys olivaceus) Immunoglobulin

Persistence of herpesvirus of eel Herpesvirus anguillae in farmed European eel Anguilla anguilla

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