Abstract
Background
Cronobacter spp. is a newly emerging pathogen that causes meningitis in infants and other diseases in elderly and immunocompromised individuals. This study was undertaken to investigate surface antigenic determinants in Cronobacter spp. using monoclonal antibodies (MAbs) and MALDI-TOF Mass spectrometry.
Results
Spleenocytes from mice that were immunized with heat-killed (20 min, 80°C) Cronobacter cells were fused with SP2 myeloma cells. Five desirable MAbs (A1, B5, 2C2, C5 and A4) were selected. MAbs A1, B5, 2C2 and C5 were of IgG2a isotype while A4 was an IgM. Specificity of the MAbs was determined by using immunoblotting with outer membrane protein preparations (OMPs) extracted from 12 Cronobacter and 6 non-Cronobacter bacteria. All MAbs recognized proteins with molecular weight ranging between 36 and 49 kDa except for one isolate (44) in which no OMPs were detected. In addition, MAbs recognized two bands (38-41 kDa) in four of the non-Cronobacterbacteria. Most of the proteins recognized by the MAbs were identified by MALDI-TOF peptide sequencing and appeared to be heterogeneous with the identities of some of them are still unknown. All MAbs recognized the same epitope as determined by an additive Index ELISA with their epitopes appeared to be conformational rather than sequential. Further, none of the MAbs recognized purified LPS from Cronobacter spp. Specificity of the MAbs toward OMPs was further confirmed by transmission electron microscopy.
Conclusions
Results obtained in this study highlight the immunological cross-reactivity among CronobacterOMPs and their Enterobacteriaceae counterparts. Nevertheless, the identity of the identified proteins appeared to be different as inferred from the MALDI-TOF sequencing and identification.
Date of this Version
6-25-2011
DOI
10.1186/1471-2180-11-148
Recommended Citation
Jaradat, Ziad W.; Rashdan, Abrar M.; Ababneh, Qotaiba O.; Jaradat, Saied A.; and Bhunia, Arun K., "Characterization of Surface Proteins of Cronobacter Muytjensii Using Monoclonal Antibodies and MALDI-TOF Mass Spectrometry" (2011). Department of Food Science Faculty Publications. Paper 4.
http://dx.doi.org/10.1186/1471-2180-11-148
Comments
This is the publisher pdf of Ziad W Jaradat, Abrar M Rashdan, Qotaiba O Ababneh, Saied A Jaradat, Arun K Bhunia. Characterization of Surface Proteins of Cronobacter Muytjensii Using Monoclonal Antibodies and MALDI-TOF Mass Spectrometry. BMC Microbiology 2011, 11:148 (25 June 2011) and is available at: 10.1186/1471-2180-11-148.