Identification of Pseudomonas aeruginosa using functional magnetic nanoparticle-based affinity capture combined with MALDI MS analysis.
PA-IL is a galactophilic lectin that is found on the outer membrane of Pseudomonas aeruginosa. Pigeon ovalbumin (POA), a phosphoprotein, contains high levels of terminal Gal alpha(1-->4)Gal units. Thus, magnetic nanoparticles with immobilized POA can be used as affinity probes for P. aeruginosa, functioning via the recognition of galactophilic PA-IL. We fabricated POA-bound nanoparticles (NPs) by immobilizing POA onto the surface of core/shell magnetic iron oxide/alumina NPs via metal-phosphate chelation. We then used the generated NPs to probe target bacteria from complex samples. The trapped bacterial cells were characterized based on their mass peak profiles obtained from MALDI MS analyses. In addition, we confirmed the determination of P. aeruginosa using a proteomic strategy: combining the resultant MALDI MS/MS spectra of its tryptic digest with protein database searching. The feasibility of using this approach to rapidly characterize P. aeruginosa from clinical samples without the need to perform culturing steps was also demonstrated.
Liu
Jr-Chi
JC
Department of Applied Chemistry, National Chiao Tung University, Hsinchu 300, Taiwan.
Chen
Wei-Jen
WJ
Li
Chen-Wei
CW
Mong
Kwok-Kong Tony
KK
Tsai
Pei-Jane
PJ
Tsai
Te-Lung
TL
Lee
Yuan C
YC
Chen
Yu-Chie
YC
D000345:Affinity Labels / Q000737:chemistry* / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D008280:Magnetics; D008872:Microwaves; D008969:Molecular Sequence Data; D053758:Nanoparticles / Q000737:chemistry*; D010047:Ovalbumin / Q000737:chemistry / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000302:isolation & purification* / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000378:metabolism
D000345:Affinity Labels; D001426:Bacterial Proteins; D055767:Immobilized Proteins; D010047:Ovalbumin; D014357:Trypsin