Production of Monoclonal Antibodies Against the Outer Cell Wall of<i>Clostridium tyrobutyricum</i>

Talbot, F.; Robreau, Georges; Gueguen, Françoise; Malcoste, R.

doi:10.1089/hyb.1994.13.45

Cited by 12 publications

(11 citation statements)

References 14 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The flagella of C. tyrobutyricum are of special interest for two reasons: first, they seem to be a major antigen in the Clostridia cell wall (28); and second, the flagellar filament protein of C. tyrobutyricum is a glycoprotein, a rare post-translational modification in Gram-positive Eubacteria (16). In this study, we have cloned amplified PCR products, and genetically characterized the coding region of the C. tyrobutyricum flagellin gene.…”

Section: Discussionmentioning

confidence: 99%

Cloning and Sequencing of the Central Region of the Flagellin Gene from the Gram‐Positive Bacterium Clostridium tyrobutyricum ATCC 25755

Arnold

Bédouet

Batina

et al. 1999

Microbiology and Immunology

Self Cite

View full text Add to dashboard Cite

The purpose of this study was to sequence the central part of the coding region of the Clostridium tyrobutyricum fiagellin gene to improve the immunoenzymatic counting of cells after milk filtration. The coding region was amplified by PCR, and the amplified products were cloned. A DNA sequence analysis of positive clones gave us 1,131 nucleotides with a partial calculated flagellin molecular mass of 40,143 Da. The flagellar filament protein sequence exhibited high levels of homology to sequences of flagellin protein from other bacteria in both N- and C-terminal parts, but little homology in the central domain. A PCR-restriction fragment length polymorphism analysis of amplified C. tyrobutyricum flagellin gene products confirmed the variability of the central domain. The flagellin mRNA was determined to be 1.1 kb in size, which suggests a monocistronic mRNA. Furthermore, the deduced protein flagellin contains eleven potential N-glycosylation sites and one sequence rich in serine, which could be modified by O-glycosylation.

show abstract

Section: Discussionmentioning

confidence: 99%

Cloning and Sequencing of the Central Region of the Flagellin Gene from the Gram‐Positive Bacterium Clostridium tyrobutyricum ATCC 25755

Arnold

Bédouet

Batina

et al. 1999

Microbiology and Immunology

Self Cite

View full text Add to dashboard Cite

show abstract

“…Milk containing spores of C. tyrobutyricum is safe for consumption as liquid milk and is suitable to be manufactured into most dairy products. However, contaminated milk even with very low numbers of spores, of about 500 spores per liter, can cause serious defects during cheese maturation [5]. Thus, the existence of a fast (<24 h), specific, and extremely sensitive method to detect this bacterium before milk processing would be advantageous, specially to screen milk that is going to be used in the production of mature cheese, thus avoiding future problems.…”

Section: Introductionmentioning

confidence: 99%

Specific peptides as alternative to antibody ligands for biomagnetic separation of Clostridium tyrobutyricum spores

et al. 2011

View full text Add to dashboard Cite

Nowadays, the reference method for the detection of Clostridium tyrobutyricum in milk is the most-probable-number method, a very time-consuming and non-specific method. In this work, the suitability of the use of superparamagnetic beads coated with specific antibodies and peptides for bioseparation and concentration of spores of C. tyrobutyricum has been assessed. Peptide or antibody functionalized nanoparticles were able to specifically bind C. tyrobutyricum spores and concentrate them up to detectable levels. Moreover, several factors, such as particle size (200 nm and 1 μm), particle derivatization (aminated and carboxylated beads), coating method, and type of ligand have been studied in order to establish the most appropriate conditions for spore separation. Results show that concentration of spore is favored by a smaller bead size due to the wider surface of interaction in relation to particle volume. Antibody orientation, related to the binding method, is also critical in spore recovery. However, specific peptides seem to be a better ligand than antibodies, not only due to the higher recovery ratio of spores obtained but also due to the prolonged stability over time, allowing an optimal recovery of spores up to 3 weeks after bead coating. These results demonstrate that specific peptides bound to magnetic nanoparticles can be used instead of traditional antibodies to specifically bind C. tyrobutyricum spores being a potential basis for a rapid method to detect this bacterial target.

show abstract

“…Consequently, the 'ideal' method to detect this bacterium should be fast (B24 h), specific and extremely sensitive because even when C. tyrobutyricum is present in very low numbers in milk (i.e. 500 spores/l), it could cause serious defects in cheese maturation (Talbot, Robreau, Gueguen, & Malcoste, 1994).…”

mentioning

confidence: 99%

Production of polyclonal antibodies against spores ofClostridium tyrobutyricum, a contaminant affecting the quality of cheese: characterisation of the immunodominant protein

Lavilla

Luis

Conesa

et al. 2008

Food and Agricultural Immunology

View full text Add to dashboard Cite

The effect of different treatments in the production of polyclonal antibodies against Clostridium tyrobutyricum spores and its immunodetection was studied. Moreover, the main protein that causes the immunological response has been characterised. Antisera from rabbits immunised with non-treated, heated or sonicated spores were tested by ELISA. Heated and sonicated spores gave a faster increase in antibody titre than non-treated spores a long time after immunisation. Heat treatment improved reactivity when it was applied to the spores prior to immunodetection. However, when the highest titre was reached, the treatment effect was not observed either in the immune response or in the immunodetection. The immunodominant protein was identified in a spore extract by transferblotting, analysed by on-line liquid chromatography tandem mass spectrometry, and characterised by comparison of its peptide fragmentation spectrum with the NCBI database. Five proteins from the family of chaperonins were shown related to the immunodominant protein of C. tyrobutyricum spore extract.

show abstract

Production of Monoclonal Antibodies Against the Outer Cell Wall ofClostridium tyrobutyricum

Cited by 12 publications

References 14 publications

Cloning and Sequencing of the Central Region of the Flagellin Gene from the Gram‐Positive Bacterium Clostridium tyrobutyricum ATCC 25755

Cloning and Sequencing of the Central Region of the Flagellin Gene from the Gram‐Positive Bacterium Clostridium tyrobutyricum ATCC 25755

Specific peptides as alternative to antibody ligands for biomagnetic separation of Clostridium tyrobutyricum spores

Production of polyclonal antibodies against spores ofClostridium tyrobutyricum, a contaminant affecting the quality of cheese: characterisation of the immunodominant protein

Contact Info

Product

Resources

About