Nano-structured support materials, their characterisation and serum protein profiling through MALDI/TOF-MS

Abstract

In the bioanalytical era, novel nano-materials for the selective extraction, pre-concentration and purification of biomolecules prior to analysis are vital. Their application as affinity binding in this regard is needed to be authentic. We report here the comparative application of derivatised materials and surfaces on the basis of nano-crystalline diamond, carbon nanotubes and fullerenes for the analysis of marker peptides and proteins by material enhanced laser desorption ionisation mass spectrometry MELDI-MS. In this particular work, the emphasis is placed on the derivatization, termed as immobilised metal affinity chromatography (IMAC), with three different support materials, to show the effectiveness of MELDI technique. For the physicochemical characterisation of the phases, near infrared reflectance spectroscopy (NIRS) is used, which is a well-established method within the analytical chemistry, covering a wide range of applications. NIRS enables differentiation between silica materials and different fullerenes derivatives, in a 3-dimensional factor-plot, depending on their derivatizations and physical characteristics. The method offers a physicochemical quantitative description in the nano-scale level of particle size, specific surface area, pore diameter, pore porosity, pore volume and total porosity with high linearity and improved precision. The measurement takes only a few seconds while high sample throughput is guaranteed.

Cromatografia de afinidade por íons metálicos imobilizados (IMAC) de biomoléculas: aspectos fundamentais e aplicações tecnológicas

“…[40][41][42][43][44][45][46] Outros materiais, como capilares monolíticos de sílica, nanotubos de carbono e fulerenos também são utilizados para a imobilização de íons metálicos na presença ou ausência de agentes quelantes. 47 Adsorventes IMAC (matriz com agente quelante imobilizado) podem ser também adquiridos comercialmente. As empresas disponibilizam adsorventes para uso em equipamentos cromatográficos de baixa pressão como, por exemplo, géis de agarose com IDA ou NTA imobilizado (GE Healthcare ® , Pierce ® , Sigma-Aldrich ® e Qiagen ® ) e, também, adsorventes para uso em equipamentos cromatográficos de média e alta pressão como, por exemplo, matrizes de sílica ou agarose altamente reticulada com IDA ou NTA imobilizados (GE Healthcare ® , Qiagen ® e Clontech Laboratoires ® ).…”

Sample preparation and fractionation for proteome analysis and cancer biomarker discovery by mass spectrometry

“…MALDI profiling, although has given better results than SELDI, still needs further standardization and evaluation [233]. Although, a recently developed carrier-based approach called MELDI is an improvement on the MALDI approach, and appears to enhance the robustness and throughput of large-scale proteomic studies [234,235], neverthless, it still needs confirmation [230].…”

Current Advances in Antibody Immobilization on Different Surfaces and Beads

“…This technique enables a prompt improvement of materials anti-human IgG and antihaemoglobin Pan et al, 2004 properties. Heigl et al, 2007;Heigl et al, 2006;Vallant et al, 2007;Najam-ul-Haq et al, 2007). …”