Production of Monoclonal Antibodies in a Mouse Model via Lipopolysaccharide Conjugates with Synthetic Polymers Specific to<i>Salmonella</i>Enteritidis O Antigen

Nalbantsoy, Ayşe; Karaboz, İsmail; Gürhan, İsmet Deliloğlu

doi:10.1089/fpd.2010.0612

Cited by 3 publications

(2 citation statements)

References 44 publications

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“…With a modified protocol based on a suitable adjuvant, immunisation with LPS could possibly have clinical applications. Results from our continuing studies shows that the LPS conjugated synthetic polymers induced higher antibody response than the naked LPS in mice (Nalbantsoy et al, 2010b). Nevertheless the obtained data needs to be confirmed by field studies.…”

Section: Discussionmentioning

confidence: 77%

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Determination ofin vivotoxicity andin vitrocytotoxicity of lipopolysaccharide isolated fromSalmonellaEnteritidis and its potential use for production of polyclonal antibody

Nalbantsoy

Karabay-Yavasoglu

Deliloglu-Gurhan

2011

Food and Agricultural Immunology

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TurkeyIn the present study, the effects of lipopolysaccharide (LPS) isolated from Salmonella Enteritidis were investigated by in vivo and in vitro methods. The mean lethal dose of 50% death (LD 50 ) was determined by a Probit test as 450-mg/mouse after 72 h post-treatment of LPS. MadinÁDarby bovine kidney (MDBK), baby hamster kidney (BHK), mouse fibroblastic cell-lines (L929), and hybridoma cell lines were treated with different amounts of LPS (0Á100 mg/ml) cultivated for 72 h in 24 well tissue culture plates. Morphological investigation was done with inverted microscope after Giemsa staining. The results suggested that MDBK, BHK, and L929 cell lines were resistant to LPS cytotoxicity due to lack of the specific membrane receptor unlike hybridoma cells. After immunisation, one priming and seven booster's diluted sera (1:100) antibody level was found to be 2.00 9 0.06Á2.41 9 0.07 optical density (OD) by enzyme linked immunosorbent assay (ELISA) on day 142. This result indicated that the LPS, instead of whole bacteria, have potential application in the immunisation.

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Section: Discussionmentioning

confidence: 77%

“…Hybridoma cell line (5A8) was obtained from a previous study (Nalbantsoy, Karaboz, & Deliloglu-Gurhan, 2010b). In brief, spleen cells from immunised Balb/c mice were fused with non-immunoglobulin producing x63-Ag8-653 murine myeloma cells purchased from the HUKUK.…”

Section: Cell Culturementioning

confidence: 99%

Determination ofin vivotoxicity andin vitrocytotoxicity of lipopolysaccharide isolated fromSalmonellaEnteritidis and its potential use for production of polyclonal antibody

Nalbantsoy

Karabay-Yavasoglu

Deliloglu-Gurhan

2011

Food and Agricultural Immunology

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Effect of Operating Conditions in Production of Diagnostic Salmonella Enteritidis O-Antigen-Specific Monoclonal Antibody in Different Bioreactor Systems

Ayyildiz-Tamis

Nalbantsoy

Elibol

et al. 2013

Appl Biochem Biotechnol

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In this study, different cultivation systems such as roller bottles (RB), 5-L stirred-tank bioreactor (STR), and disposable bioreactors were used to cultivate hybridoma for lab-scale production of Salmonella Enteritidis O-antigen-specific monoclonal antibody (MAb). Hybridoma cell line was cultivated in either serum-containing or serum-free medium (SFM) culture conditions. In STR, MAb production scaled up to 4 L, and production capabilities of the cells were also evaluated in different featured production systems. Moreover, the growth parameters of the cells in all production systems such as glucose consumption, lactate and ammonia production, and also MAb productivities were determined. Collected supernatants from the reactors were concentrated by a cross-flow filtration system. In conclusion, cells were not adapted to SFM in RB and STR. Therefore, less MAb titer in both STR and RB systems with SFM was observed compared to the cultures containing fetal bovine serum-supplemented medium. A higher MAb titer was gained in the membrane-aerated system compared to those in STR and RB. Although the highest MAb titer was obtained in the static membrane bioreactor system, the highest productivity was obtained in STR operated in semicontinuous mode with overlay aeration.

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Metabolic Activity and Monoclonal Antibody Production of Salmonella Enteritidis O and H Antigen Specific Hybridoma Cells in Static Culture

2011

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The aim of the present work was to study the kinetics of two hybridomas that produce monoclonal antibody against Salmonella Enteritidis O (5A8) and H (D7) antigen. The hybridomas originated from the Ag8x653 (5A8) and Sp2/O (D7) myeloma cell lines. The relationship between the uptake of glucose and glutamine and the release of the lactate and ammonia and monoclonal antibody production into hybridoma growth were investigated in static culture with serum-containing DMEM/F:12 medium for the determination of pilot-production strategies of the hybridomas. Results showed that glucose and glutamine concentrations were reduced, with an increase in ammonia and lactate concentration in the culture medium. The hybridoma cell line 5A8 has shown lower metabolic activities compared with D7, whereas its monoclonal antibody productivity was found to be two-fold higher than the D7. MAb production by the hybridoma cell line 5A8 seems promising, considering the moderate level of productivity compared to that found in the literature.

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Production of Monoclonal Antibodies in a Mouse Model via Lipopolysaccharide Conjugates with Synthetic Polymers Specific toSalmonellaEnteritidis O Antigen

Cited by 3 publications

References 44 publications

Determination ofin vivotoxicity andin vitrocytotoxicity of lipopolysaccharide isolated fromSalmonellaEnteritidis and its potential use for production of polyclonal antibody

Determination ofin vivotoxicity andin vitrocytotoxicity of lipopolysaccharide isolated fromSalmonellaEnteritidis and its potential use for production of polyclonal antibody

Effect of Operating Conditions in Production of Diagnostic Salmonella Enteritidis O-Antigen-Specific Monoclonal Antibody in Different Bioreactor Systems

Metabolic Activity and Monoclonal Antibody Production of Salmonella Enteritidis O and H Antigen Specific Hybridoma Cells in Static Culture

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