Tuberc Respir Dis.  2006 Apr;60(4):412-418. 10.4046/trd.2006.60.4.412.

Development of Protein Chip for Diagnosis of Chlamydophia Pneumoniae

• Kim WJ
• Lee HY
• Lee SJ
• Jung SH
• Yuk JS
• Ha KS
• Jung KS

• KMID: 1719366
• DOI: http://doi.org/10.4046/trd.2006.60.4.412

Abstract

BACKGROUND: The diagnosis of chlamydial infection is based on serology. The current gold standard of diagnosis is MIF(microimmunofluorescence), but this modality is subjective and time-consuming. Protein microarray with using a SPR(surface plasmon resonance) sensor has recently been suggested as a method for detecting infection. For developing a protein chip to diagnose chlamydial infection, EBs(elementary bodies) were immobilized on a gold chip and the interaction between an antibody for Chlamydophila pneumoniae and the EBs(elementary bodies) immobilized on the surface of the gold chip was measured by using an SPR sensor.
METHODS
For the surface antigen, the EBs of Chlamydophila pneumoniae LKK1 were purified. Charged arrays were prepared by using PDDA(polydiallyldimethylammonium chloride) which has a positive charge. After immobilization of the chlamydial EBs on the PDDA surface, the investigation of the surface was done with using atomic force microscopy. After the antibody for C. pneumoniae was applied on chip, we monitored the SPR wavelength-shift to detect any antigen-antibody interaction with using a self-assembled SPR sensor.
RESULTS
The chlamydial EBs on the positively charged PDDA were visible on the surface with using atomic force microscopy. The SPR wavelength increased after interaction of antibody for C. pneumoniae with the EBs immobilized on charged gold surface. The wavelength-shift was correlated with the concentration of antigens.
CONCLUSION
The surface immobilization of EBs on the gold surface with the charged arrays was identified and the antigen-antibody interaction on the gold chip was detected via the SPR sensor. Further investigations are needed to apply this technique to the clinical field.

Keyword

Chlamydophila pneumoniae; Protein array analysis

MeSH Terms

Antigens, Surface
Chlamydial Pneumonia
Chlamydophila pneumoniae
Diagnosis*
Immobilization
Microscopy, Atomic Force
Pneumonia*
Protein Array Analysis*
Antigens, Surface

Figure

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  Figure 1 Scheme of self-assembled SPR sensor.

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  Figure 2 Atomic force microscopy image of C. pneumoniae(EB) on PDDA (polydiallyldimethylammonium chloride) gold surface

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  Figure 3 Scanning electron microscopy image of C. pneumoniae(EB) on PDDA (polydiallyldimethylammonium chloride) gold surface

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  Figure 4 SPR(surface plasmon resonance) wavelength change after antibody-antigen interaction

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