Immunomagnetic separation and MS/SPR end-detection combined procedure for rapid detection of Staphylococcus aureus and protein A.
The aim of this study was to establish an IMS-MS/SPR technique for the detection of Staphylococcus aureus (S. aureus) and Staphylococcus protein A (SPA) at the same time, which consists of isolating S. aureus and trapping-enrichmenting its SPA by IMS, and the end point is determined by using either MS or SPR measurements. Magnetic bead (MB) containing aldehyde group was synthesized with latex-polymerization and immunomagnetic bead (IMB) was fabricated by modifying its surface with an oriented layer of human IgG in covalent linkage. As soon as sample of pulverator-treated bacterial cell lysate (10(8) cfu/mL) was incubated with IMB at 4 degrees C for 30 min, SPA was captured and separated from the mixed solution in a few minutes by the IMB and then detected with mass spectrometry after washing. SPR was used to detect S. aureus quantitatively in situ at the end-detection procedure. All in all, this technique can be employed to detect rapidly SPA and S. aureus within 2h and also be applied to detect other cells or their membrane proteins with changed modified antibodies.
Chen
Lingli
L
Department of Microbiology, Hunan Normal University, Changsha, Hunan 410081, People's Republic of China.
Deng
Le
L
Liu
Linlin
L
Peng
Zhihui
Z
D000447:Aldehydes; D018189:Immunomagnetic Separation; D013058:Mass Spectrometry; D008863:Microspheres; D013205:Staphylococcal Protein A / Q000032:analysis* / Q000737:chemistry; D013211:Staphylococcus aureus / Q000737:chemistry / Q000302:isolation & purification*; D020349:Surface Plasmon Resonance
D000447:Aldehydes; D013205:Staphylococcal Protein A