Murine monoclonal antibodies against staphylococcal enterotoxin B: production and characterization

Goyache, J.; Orden, José A.; Blanco, José Luis Santiago; Hernández, Javier; Doménech, Ana; Suárez, G.; Gómez-Lucı́a, Esperanza

doi:10.1111/j.1574-6968.1992.tb05002.x

Cited by 7 publications

(5 citation statements)

References 26 publications

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“…Female BALB/c mice (4–6‐week‐old) were inoculated with the different BLV preparations using either the intrasplenic protocol ( Spitz et al, 1984) or the conventional intraperitoneal immunization protocol ( Goyache et al, 1992). Fusions between P3/NS1/1/Ag 4 myeloma and spleen cells from hyperimmunized mice were performed with PEG 1540 (Sigma, St Louis, MO).…”

Section: Methodsmentioning

confidence: 99%

Production and Characterization of Monoclonal Antibodies against Bovine Leukaemia Virus using Various Crude Antigen Preparations: a Comparative Study

Llames

Gómez-Lucı́a

Doménech

et al. 2000

Journal of Veterinary Medicine, Series B

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A total of 59 monoclonal antibodies (mAbs) specific against the bovine leukaemia virus (BLV) using different antigen preparations was produced. The five antigen preparations for immunizing BALB/c mice were: live cells (CEL), sonicated and ultracentrifuged cells (SOC), cell lysates (LYS), semi-purified BLV (PV), and formalin-treated cells (FOR) from two cell lines permanently infected with BLV (FLK-BLV and BLV-bat2). These viral component presentations were selected to obtain mAbs against specific BLV proteins: located on the cell surface (FOR and CEL), in free virus particles (PV) and intracellular viral proteins (SOC and LYS). Two antigen preparations (SOC and LYS) were lethal to the mice following the intravenous and intrasplenic routes. Six fusions were performed in this study that rendered specific antibodies against BLV. The highest number of hybridomas was produced with SOC; however, the majority of the hybridomas produced (> 90%) were against cellular proteins. Even though immunization with PV gave the lowest number of hybridomas, the majority of them were specific against BLV. Based on the reactivity of the mAbs in Western blot (WB), we classified the mAbs into five groups, namely anti-gp51SU (39 mAbs), anti-gp30TM (six mAbs), anti-Pr72env (nine mAbs), anti-Pr66gag-pro (one mAb) and anti-Prgag (four mAbs). A very high percentage of the mAbs produced (48 of 59) reacted with gp51SU, suggesting that this is the most immunogenic and accessible BLV protein presented in the different antigen preparations. The majority of our mAbs recognized more than one band in WB, suggesting that, aside from reacting with mature proteins, the mAbs also recognized viral precursors.

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Section: Methodsmentioning

confidence: 99%

Production and Characterization of Monoclonal Antibodies against Bovine Leukaemia Virus using Various Crude Antigen Preparations: a Comparative Study

Llames

Gómez-Lucı́a

Doménech

et al. 2000

Journal of Veterinary Medicine, Series B

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“…Chloroform was added to a final concentration of 15 % (v/v) and the mixture was stirred with a magnetic stirrer for 5 min, Results and Discussion In this study, three MAbs produced directly against SEB were analysed. All of them reacted with SEB and with its precursor, as seen by immunoblot (GOYACHE et al, 1992), but not with any of the other staphylococcal enterotoxins assayed (SEA, SEC,, SED or SEE).…”

Section: Food Extractsmentioning

confidence: 87%

“…MAbs against pure SEB were obtained using standard techniques and characterized by ELISA (GOYACHE et al, 1992). The MAbs used in this study (B5, B12 and B14) reacted specifically with SEB both in ELISA and immunoblot (GOYACHE et al, 1992).…”

Section: Monoclonal Antibodiesmentioning

confidence: 99%

“…Several papers have described the preparation of murine monoclonal antibodies (MAbs) against SEB (THOMPSON et a]., 1984(THOMPSON et a]., , 1985LAPEYRE et al, 1987;LIN et al, 1988;KIENLE and BUSCHMANN, 1989;DUPONT, et al, 1990;GOYACHE et al, 1992), but in only two cases have these MAbs been applied to assay systems for the detection of SEB in food samples using MAbs exclusively (THOMPSON et al, 1986;DUPONT et al, 1990 (LAPEYRE et al, 1987(LAPEYRE et al, , 1988) described an ELISA system which involves the use of a specific MAb and a polyclonal rabbit serum directed against the four enterotoxins A, B, C, and D, a rationale which is difficult to understand if one is interested in eliminating the need for producing polyclonal rabbit serum (THOMPSON et a]., 1985). The aim of this study is to describe an ELISA technique using a pair of murine MAbs which react specifically with SEB, and which can be used effectively for the detection of SEB from food extracts.…”

Section: Introductionmentioning

confidence: 99%

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Double‐sandwich Enzyme‐linked Immunosorbent Assay (ELISA) with Murine Monoclonal Antibodies for Detection of Staphylococcal Enterotoxin B

Goyache

Orden

Blanco

et al. 1994

Journal of Veterinary Medicine, Series B

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Sum m a r yThree monoclonal antibodies (MAbs) which react specifically with staphylococcal enterotoxin B (SEB) were studied for their suitability for use in ELISA. One of them (MAb B14) worked well as a coating antibody; MAb B12 was shown to be a good probing antibody (conjugated with peroxidase) when ELISA plates were coated with MAb B14. This effective pair of MAbs (B14-B12P0) is able to detect 0.625 ng SEB ml, and to distinguish between SEB and other proteins present in food extracts.

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“…Combination therapy using cocktails of monoclonal antibodies (mAbs) may be necessary to cover, for example, toxin heterogeneity 82 . Passive administration of mAbs for toxins such as ricin 83 and staphylococcal enterotoxin has been shown to protect mice from lethal toxin challenge 84 . Several reports have suggested that therapeutic antibodies can target both intracellular and extracellular pathogens 85 , 86 .…”

Section: Passive Immune Therapies: a Feasible Option?mentioning

confidence: 99%

Novel strategies for controlling Streptococcus pyogenes infection and associated diseases: from potential peptide vaccines to antibody immunotherapy

2009

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Infections caused by group A streptococcus (GAS) represent a public health problem in both developing and developed countries. The current available methods of prevention are either inadequate or ineffective, which is highlighted by the resurgence in invasive GAS infections over the past two decades. The management of GAS and associated diseases requires new and improved approaches. This review discusses various potential approaches in controlling GAS infections, ranging from prophylactic vaccines to antibody immunotherapy.

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Murine monoclonal antibodies against staphylococcal enterotoxin B: production and characterization

Cited by 7 publications

References 26 publications

Production and Characterization of Monoclonal Antibodies against Bovine Leukaemia Virus using Various Crude Antigen Preparations: a Comparative Study

Production and Characterization of Monoclonal Antibodies against Bovine Leukaemia Virus using Various Crude Antigen Preparations: a Comparative Study

Double‐sandwich Enzyme‐linked Immunosorbent Assay (ELISA) with Murine Monoclonal Antibodies for Detection of Staphylococcal Enterotoxin B

Novel strategies for controlling Streptococcus pyogenes infection and associated diseases: from potential peptide vaccines to antibody immunotherapy

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