In:
PROTEOMICS – Clinical Applications, Wiley, Vol. 7, No. 5-6 ( 2013-06), p. 378-383
Abstract:
We aim to develop a protein microarray platform capable of presenting both natural and denatured forms of proteins for antibody biomarker discovery. We will further optimize plasma screening protocols to improve detection. Experimental design We developed a new covalent capture protein microarray chemistry using H alo T ag fusion proteins and ligand. To enhance protein yield, we used H e L a cell lysate as an in vitro transcription translation ( IVTT ) system. E scherichia coli lysates were added to the plasma blocking buffer to reduce nonspecific background. These protein microarrays were probed with plasma samples and autoantibody responses were quantified and compared with or without denaturing buffer treatment. Results We demonstrated that protein microarrays using the covalent attachment chemistry endured denaturing conditions. Blocking with E . coli lysates greatly reduced the background signals and expression with IVTT based on H e L a cell lysates significantly improved the antibody signals on protein microarrays probed with plasma samples. Plasma samples probed on denatured protein arrays produced autoantibody profiles distinct from those probed on natively displayed proteins. Conclusions and clinical relevance This versatile protein microarray platform allows the display of both natural and denatured proteins, offers a new dimension to search for disease‐specific antibodies, broadens the repertoire of potential biomarkers, and will potentially yield clinical diagnostics with greater performance.
Type of Medium:
Online Resource
ISSN:
1862-8346
,
1862-8354
DOI:
10.1002/prca.201200062
Language:
English
Publisher:
Wiley
Publication Date:
2013
detail.hit.zdb_id:
2317130-3
