This is a protocol preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Method Article
Sandeep Kumar Vashist
HSG-IMIT - Institut für Mikro- und Informationstechnik; Laboratory for MEMS Applications, Department of Microsystems Engineering -IMTEK, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany
Corresponding Author
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
A rapid and highly-sensitive procedure has been developed for surface plasmon resonance (SPR)-based immunoassay (IA) for human fetuin A (HFA). It employs a highly-simplified antibody (Ab) immobilization strategy, which only involves sequentially the dispensing of anti-HFA capture Ab diluted in 1% (v/v) 3-aminopropyltriethoxysilane (APTES) onto a potassium hydroxide (KOH)-treated gold (Au)-coated SPR chip followed by its incubation for 30 min. The devised Ab immobilization strategy enables the leach-proof binding of capture Ab, resulting in highly reproducible and cost-effective HFA IAs. The SPR IA detected HFA with a dynamic range, limit of detection, and analytical sensitivity of 0.3-20 ng mL-1, 0.7 ng mL-1 and 1 ng mL-1, respectively. The detecting chip can be easily regenerated after each analyis by treatment with 10 mM glycine-HCl, pH 2.0. The Ab-bound SPR chip can be stored at 4°C for up to 4 months without any significant loss of its functional activity. The developed IA procedure has been employed to detect HFA spiked in diluted human whole blood and plasma. Moreover, the developed SPR IA has high analytical precision as its results are correlated well with those of conventional HFA sandwich ELISA.
Keywords
Surface plasmon resonance, immunoassay, one-step antibody immobilization, 3-aminopropyltriethoxysilane, human fetuin A
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
The authors declare no competing financial interests.
Loading...
Version History
CURRENT STATUS: Posted
Version 1
Posted 30 Apr, 2014
No community comments so far
Metrics
Comments: 0
HTML Views: ...
Associated Publications
Surface plasmon resonance-based immunoassay for human fetuin A
by S. K. Vashist, E. M. Schneider, and J. H. T. Luong
The Analyst (29 April, 2014)
Method Article
Sandeep Kumar Vashist
HSG-IMIT - Institut für Mikro- und Informationstechnik; Laboratory for MEMS Applications, Department of Microsystems Engineering -IMTEK, University of Freiburg, Georges-Koehler-Allee 103, 79110 Freiburg, Germany
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 30 Apr, 2014
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY-NC 3.0 License. Read Full License
A rapid and highly-sensitive procedure has been developed for surface plasmon resonance (SPR)-based immunoassay (IA) for human fetuin A (HFA). It employs a highly-simplified antibody (Ab) immobilization strategy, which only involves sequentially the dispensing of anti-HFA capture Ab diluted in 1% (v/v) 3-aminopropyltriethoxysilane (APTES) onto a potassium hydroxide (KOH)-treated gold (Au)-coated SPR chip followed by its incubation for 30 min. The devised Ab immobilization strategy enables the leach-proof binding of capture Ab, resulting in highly reproducible and cost-effective HFA IAs. The SPR IA detected HFA with a dynamic range, limit of detection, and analytical sensitivity of 0.3-20 ng mL-1, 0.7 ng mL-1 and 1 ng mL-1, respectively. The detecting chip can be easily regenerated after each analyis by treatment with 10 mM glycine-HCl, pH 2.0. The Ab-bound SPR chip can be stored at 4°C for up to 4 months without any significant loss of its functional activity. The developed IA procedure has been employed to detect HFA spiked in diluted human whole blood and plasma. Moreover, the developed SPR IA has high analytical precision as its results are correlated well with those of conventional HFA sandwich ELISA.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Loading...
Associated Publications
Surface plasmon resonance-based immunoassay for human fetuin A
by S. K. Vashist, E. M. Schneider, and J. H. T. Luong
The Analyst (29 April, 2014)