Overview on modern approaches to speed up protein identification workflows relying on enzymatic cleavage and mass spectrometry-based techniques

• Published in: Analytica chimica acta 2009-09, Vol.650 (2), p.151-159
• Main Authors: Capelo, J.L., Carreira, R., Diniz, M., Fernandes, L., Galesio, M., Lodeiro, C., Santos, H.M., Vale, G.
• Format: Article
• Language: English
• Subjects: Analytical biochemistry: general aspects, technics, instrumentation; Analytical chemistry; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chemistry; Enzymes - metabolism; Exact sciences and technology; Fast methods; Fundamental and applied biological sciences. Psychology; Mass spectrometry; Mass Spectrometry - methods; Protein Denaturation; Protein identification; Protein Processing, Post-Translational; Proteins - analysis; Proteins - chemistry; Proteomics; Spectrometric and optical methods
• ISSN: 0003-2670; 1873-4324
• DOI: 10.1016/j.aca.2009.07.034

Recent tools addressed to accelerate the different steps of the sample treatment for protein identification in modern workflows are reviewed and critically commented in this manuscript. Heating, microspin columns, ultrasonic energy, high pressure, infrared energy, microwave energy, alternating electric fields and microreactors are outlined as useful tools that can be used to accelerate all or some of the following steps for in-gel or in-liquid based approaches for protein identification: (i) protein dissolution/denaturation, (ii) protein reduction, (iii) protein alkylation and (iv) protein digestion. The advantages and drawbacks, along with the main differences among the different tools are also commented. Future prospects for hyphenation of methods are also discussed. Researchers are informed also in this work regarding the main problems to be found when implementing any of the above mentioned methods.