Production and characterization of anti-bifidobacteria monoclonal antibodies and their application in the development of an immuno-culture detection method

Amrouche, Tahar; Boutin, Yvan; Moroni, Olivier; Kheadr, Ehab; Fliss, Ismaı̈l

doi:10.1016/j.mimet.2005.07.009

Cited by 8 publications

(6 citation statements)

References 30 publications

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“…Another study also reported the development of anti‐bifidobacteria mAbs used for immuno‐culture methods (Amrouche et al . ). However, mAbs production is more expensive and time consuming than pAbs.…”

Section: Discussionmentioning

confidence: 97%

“…A few studies reported the mAbs applied to LAB and probiotics, such as Stuknyte and colleagues who developed a mAb targeting L. helveticus SLP (from a different strain) for its detection in cheese by western blot (Stuknyte et al 2014). Another study also reported the development of anti-bifidobacteria mAbs used for immuno-culture methods (Amrouche et al 2006). However, mAbs production is more expensive and time consuming than pAbs.…”

Section: Antibody Productionmentioning

confidence: 99%

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Flow cytometry: a versatile technology for specific quantification and viability assessment of micro-organisms in multistrain probiotic products

2018

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Aims: Classical microbiology techniques are the gold standard for probiotic enumeration. However, these techniques are limited by parameters of time, specificity and incapacity to detect viable but nonculturable (VBNC) microorganisms and nonviable cells. The aim of the study was to evaluate flow cytometry as a novel method for the specific quantification of viable and nonviable probiotics in multistrain products. Methods and Results: Custom polyclonal antibodies were produced against five probiotic strains from different species (Bifidobacterium bifidum R0071, Bifidobacterium longum ssp. infantis R0033, Bifidobacterium longum ssp. longum R0175, Lactobacillus helveticus R0052 and Lactobacillus rhamnosus R0011). Evaluation of specificity confirmed that all antibodies were specific at least at the subspecies level. A flow cytometry method combining specific antibodies and viability assessment with SYTO â 24 and propidium iodide was applied to quantify these strains in three commercial products. Analyses were conducted on two flow cytometry instruments by two operators and compared with classical microbiology using selective media. Results indicated that flow cytometry provides higher cell counts than classical microbiology (P < 0Á05) in 73% of cases highlighting the possible presence of VBNC. Equivalent performances (repeatability and reproducibility) were obtained for both methods. Conclusions: This study showed that flow cytometry methods can be applied to probiotic enumeration and viability assessment. Combination with polyclonal antibodies can achieve sufficient specificity to differentiate closely related strains. Significance and Impact of the Study: Flow cytometry provides absolute and specific quantification of viable and nonviable probiotic strains in a very short time (<2 h) compared with classical techniques (>48 h), bringing efficient tools for research and development and quality control.

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Section: Discussionmentioning

confidence: 97%

Section: Antibody Productionmentioning

confidence: 99%

Flow cytometry: a versatile technology for specific quantification and viability assessment of micro-organisms in multistrain probiotic products

2018

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“…Monoclonal antibodies could be prepared also for the assay of whole cells such as Listeria monocytogenes; Crowley et al (1999) used prepared antibodies for the construction of an amperometric immunosensor as well as ELISA. In other studies, monoclonal antibodies against the cell wall proteins of Bifidobacterium longum (Amrouche et al 2006) andB. anthracis lethal toxin (Little et al 1990) were prepared in suitable amounts and quality.…”

Section: Production Of Monoclonal Antibodiesmentioning

confidence: 99%

Monoclonal and polyclonal antibodies production - preparation of potent biorecognition element

Pohanka¹

2009

JAB

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Antibodies are macromolecular structures with a high affinity to a molecule called the antigen. Though the natural aim of antibodies is the protection of the body from pathogens, they have been found to play a useful part in a number of specific treatments and diagnoses. This wider role for antibodies extends to assays where antibodies provide a recognition capability improving available physical methods. Photometrical methods such as ELISA or flow cytometry and electrochemical, optical or piezoelectric immunosensors (biosensors) are typical examples. The present review summarizes facts about the parameters and the production of antibodies. The structure of immunoglobulins, and the production and purification of monoclonal and polyclonal antibodies are described in four chapters. The review was written in order to collect the available knowledge on antibodies into one study improving orientation in this field for anyone wanting to construct immunoassays.

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“…2 The role of physicochemical properties on immunomodulating properties of EPS has been demonstrated, and not all EPS are able to stimulate the immune system. 6 Zwitterionic EPS induce memory immune responses and can be presented in the context of MHCII or molecules such as C3. 7,8 Even though it has been demonstrated that some EPS orally or parenterally administered are able to modulate the immune response, the mechanism involved remains unknown.…”

Section: ' Introductionmentioning

confidence: 99%

Oral Administration of Kefiran Induces Changes in the Balance of Immune Cells in a Murine Model

Medrano

Racedo

Rolny

et al. 2011

J. Agric. Food Chem.

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The aim of the present study was to evaluate the effect of the oral administration of kefiran on the balance of immune cells in a murine model. Six week old BALB/c mice were treated with kefiran (300 mg/L) for 0, 2 and 7 days. Kefiran treatment increased the number of IgA+ cells in lamina propria after 2 and 7 days. Percentage of B220+/MHCII(high) cells in mesenteric lymph nodes (2 days) and Peyer's patches (7 days) was higher compared to untreated control mice. An increase of macrophages (F4/80+ cells) was observed in lamina propria and peritoneal cavity (2 and 7 days). In contrast, at day 7, macrophage population decreased in Peyer's patches. These results show the ability of kefiran to modify the balance of immune cells in intestinal mucosa. This property could be highly relevant for the comprehension of the probiotic effect attributed to kefir.

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Production and characterization of anti-bifidobacteria monoclonal antibodies and their application in the development of an immuno-culture detection method

Cited by 8 publications

References 30 publications

Flow cytometry: a versatile technology for specific quantification and viability assessment of micro-organisms in multistrain probiotic products

Flow cytometry: a versatile technology for specific quantification and viability assessment of micro-organisms in multistrain probiotic products

Monoclonal and polyclonal antibodies production - preparation of potent biorecognition element

Oral Administration of Kefiran Induces Changes in the Balance of Immune Cells in a Murine Model

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