Reproducible Two-Dimensional Capillary Electrophoresis Analysis of Barrett's Esophagus Tissues
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- 作者：James R. Kraly ; Megan R. Jones ; David G. Gomez ; Jane A. Dickerson ; Melissa M. Harwood ; Michael Eggertson ; Thomas G. Paulson ; Carissa A. Sanchez ; Robert Odze ; Ziding Feng ; Brian J. Reid ; Norman J. Dovic
- 刊名：Analytical Chemistry
- 出版年：2006
- 出版时间：September 1, 2006
- 年：2006
- 卷：78
- 期：17
- 页码：5977 - 5986
- 全文大小：1096K
- 年卷期：v.78,no.17(September 1, 2006)
- ISSN：1520-6882
文摘
We have constructed a high-speed, two-dimensional capillary electrophoresis system with a compact and high-sensitivity fluorescence detector. This instrument is usedfor the rapid and reproducible separations of Barrett'sesophagus tissue homogenates. Proteins and biogenicamines are labeled with the fluorogenic reagent 3-(2-furoyl)quinoline-2-carboxaldehyde. Labeled biomoleculesare separated sequentially in two capillaries. The firstcapillary employs capillary sieving electrophoresis usinga replaceable sieving matrix. Fractions are successivelytransferred to a second capillary where they undergoadditional separation by micellar electrokinetic capillarychromatography. The comprehensive two-dimensionalseparation requires 60 min. Within-day migration timereproducibility is better than 1% in both dimensions forthe 50 most intense features. Between-day migration timeprecision is 1.3% for CSE and better than 0.6% for MECC.Biopsies were obtained from the squamous epitheliumin the proximal tubular esophagus, Barrett's epitheliumfrom the distal esophagus, and fundus region of thestomach from each of three Barrett's esophagus patientswith informed consent. We identified 18 features from thehomogenate profiles as biogenic amines and amino acids.For each of the patients, Barrett's biopsies had more than5 times the levels of phenylalanine and alanine as compared to squamous tissues. The patient with high-gradedysplasia shows the highest concentrations for 13 of theamino acids across all tissue types. Concentrations ofglycine are 40 times higher in squamous biopsies compared to Barrett's and fundal biopsies from the patientwith high-grade dysplasia. These results suggest that two-dimensional capillary electrophoresis may be of value forthe rapid characterization of endoscopic and surgicalbiopsies.