Advanced Techniques in Clinical Practice: Use of Lab-on-a-Chip Electrophoresis and Other Methods in Protein Profiling

Abstract

IntroductionEveryday laboratory analyses are done in order to contribute to precise medical diagnosis in patients with different diseases. The techniques developed include methods which are easy to handle, provide results with satisfying selectivity and sensitivity, and are fast enough to give results in a short time. Kratak sadr`aj: U klini~koj praksi, proteini se analiziraju kao parametri va`ni za odre|ivanje i le~enje razli~itih bolesti. Postoje dva opsega analiza: analize kompletnog proteinskog profila ili odre|ivanje izolovanog proteina. U ovom radu, pored kori{}enja uobi~ajenih metoda, pre svega elektroforeze, u analizu proteina uklju~ene su nove tehnike. Laboratorija na ~ipu je elektroforetska tehnika koja, opti mizovana, omogu}ava analize ukupnog proteinskog profila. Kada se normalni uzorci uporede sa uzorcima dobijenim od pacijenata s razli~itim neurolo{kim oboljenjima, postaju uo~lji vi karakteristi~ni obrasci. Tako|e, mogu}e je uspostaviti korelaciju i pore|enje izme|u nedavno razvijenih metoda »mikro~ip« elektroforeze i rezultata dobijenih kori{}enjem uobi~ajenih tehnika. Kada je neophodna ana liza specifi~nog proteina, masena spektrometrija je dala najbolje rezultate, {to se ti~e selektivnosti kao i specifi~nosti analiza. Postoji uverenje da je cistatin C potencijalni biomarker za neurolo{ka oboljenja, stoga je razvijen metod masene spektrometrije kako bi se po stigle kvalitativne i kvantitativne analize biolo{kih te~nosti. Pomo}u metode imunoeseja masenom spektrometrijom (MSIA), cistatin C je lako izolovan i analiziran a kompletna analiza trajala je nekoliko minuta. Maseni spektri koji su nastali kao rezultat otkrili su razli~ite nivoe izoformi cistatina C u serumu i uzorcima cerebrospinalne te~nosti. Summary: Proteins in clinical practice are analyzed as important parameters in the determination and treatment of different diseases. The scopes of the analyses are mainly concentrated in two levels -analyses of the complete protein profile, or determination of an isolated protein. In this work, despite of the use of conventional methods, mainly electrophoresis, new techniques have been implemented in protein analyses. Lab-on-a-chip is an electrophoretic techni que that, when optimized, provides analyses of the total protein profile. When normal samples are compared to samples obtained from patients with different neurological diseases, characteristic patterns can be noted. Also, correlation and comparison can be made between the newly developed microchip electrophoresis method and the results obtained using the conventional techniques. When an analysis of a specific protein is necessary, mass spectrometry has proven to give best results, in both the se lectivity and specificity of analyses. It is believed that cysta tin C is a potential biomarker in neurological diseases; the refore, the mass spectrometry method has been developed in order to obtain qualitative and quantitative analyses of biological fluids. Using the developed method of mass spectro metry immunoassay (MSIA), cystatin C was easily is...

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