title: Application of capillary isoelectric focusing and peptide mass fingerprinting in carbohydrate-deficient transferrin detection.
abstract: Carbohydrate-deficient transferrin (CDT) is a specific biomarker of alcohol abuse and is widely used in clinical diagnosis to detect and follow up excessive alcohol consumption. However, false %CDT results still exist in CDT detection, because of interference from genetic variants and the lack of standardization in CDT analysis. Therefore, it is still very important to find a method with high sensitivity and high accuracy for CDT detection. Here, we compared the detection sensitivity and accuracy of pI values based methods [isoelectric focusing on polyacrylamide gel electrophoresis (IEF-PAGE) and capillary isoelectric focusing (CIEF)] with hydrophobic characteristic based methods [reversed-phase high-performance liquid chromatography (RP-HPLC)] on CDT detection. Moreover, we investigated the potential of peptide mass fingerprinting (PMF), a method based on the mass spectrometry to identify human transferrin (HTf) variants including CDT isoforms and genetic variants, based on their specific peptide masses. Results indicated that PMF can identify HTf variants including CDT isoforms and genetic variants based on their specific peptides, and CIEF showed higher sensitivity detection of HTf variants than RP-HPLC and IEF-PAGE did. Accordingly, we suggest that PMF is suitable for identifying CDT with high accuracy, and CIEF has potential for detection of CDT and genetic variants with high sensitivity; moreover, they are both worth further investigation in clinical diagnosis.
authors.affiliation: Key Laboratory of MOE for Cell Biology and Tumor Cell Engineering, School of Life Sciences, Xiamen University, Xiamen 361005, China.
authors.forename: Lian-Zhong
authors.initials: LZ
authors.lastname: Luo
authors.forename: Hong-Wei
authors.initials: HW
authors.lastname: Jin
authors.forename: He-Qing
authors.initials: HQ
authors.lastname: Huang
mesh_terms: D000437:Alcoholism / Q000378:metabolism; D015415:Biomarkers / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods*; D008297:Male; D008970:Molecular Weight; D010449:Peptide Mapping / Q000379:methods*; D020033:Protein Isoforms / Q000737:chemistry / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014168:Transferrin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D015415:Biomarkers; D020033:Protein Isoforms; D014168:Transferrin; C056759:carbohydrate-deficient transferrin