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  • Chemically modified diamond-like carbon (DLC) for protein enrichment and profiling by MALDI-MS.

Chemically modified diamond-like carbon (DLC) for protein enrichment and profiling by MALDI-MS.

Amino acids (2011-11-15)
M Najam-ul-Haq, M Rainer, C W Huck, M N Ashiq, G K Bonn
ABSTRACT

The development of new high throughput methods based on different materials with chemical modifications for protein profiling of complex mixtures leads towards biomarkers; used particularly for early diagnosis of a disease. In this work, diamond-like carbon (DLC) is developed and optimized for serum protein profiling by matrix-assisted laser/desorption ionization mass spectrometry (MALDI-MS). This study is carried out in connection with a material-based approach, termed as material-enhanced laser desorption ionization mass spectrometry. DLC is selected as carrier surface which provides large surface to volume ratio and offers high sensitivity. DLC has a dual role of working as MALDI target while acting as an interface for protein profiling by specifically binding peptides and proteins out of serum samples. Serum constituents are bound through immobilized metal ion affinity chromatography (IMAC) functionality, created through glycidyl methacrylate polymerization under ultraviolet light followed by further derivatization with iminodiacetic acid and copper ion loading. Scanning electron microscopy highlights the morphological characteristics of DLC surface. It could be demonstrated that IMAC functionalized DLC coatings represent a powerful material in trapping biomolecules for their further analysis by MALDI-MS resulting in improved sensitivity, specificity and capacity in comparison to other protein-profiling methods.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Glycidyl methacrylate, 97%, contains 100 ppm monomethyl ether hydroquinone as inhibitor
Sigma-Aldrich
Glycidyl methacrylate, ≥97.0% (GC)
Sigma-Aldrich
Iminodiacetic acid, purum, ≥98.0% (T)
Sigma-Aldrich
Iminodiacetic acid, 98%