A proof-of-concept analysis of carbohydrate-deficient transferrin by imaged capillary isoelectric focusing and in-capillary immunodetection
| Autor: | Christopher Heger, Jiaqi Wu, Annegret Boge, Charles H. Haitjema |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Chronic alcohol abuse
Capillary action Carbohydrate deficient transferrin Proof of Concept Study 01 natural sciences General Biochemistry Genetics and Molecular Biology 03 medical and health sciences medicine Humans 030304 developmental biology chemistry.chemical_classification 0303 health sciences Chromatography medicine.diagnostic_test Isoelectric focusing 010401 analytical chemistry Transferrin Serum samples Immunohistochemistry 0104 chemical sciences Isoelectric point chemistry Immunoassay Isoelectric Focusing Biotechnology |
| Zdroj: | BioTechniques. 68:85-90 |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/btn-2019-0058 |
| Popis: | Carbohydrate-deficient transferrin (CDT) is a reliable biomarker for chronic alcohol abuse. We developed a method for CDT analysis by capillary isoelectric focusing, followed by immunodetection directly in the capillary, in an automated fashion and on a single platform (Peggy Sue™; ProteinSimple, CA, USA). Transferrin glycoforms in serum samples, including disialo-transferrin, were separated and their apparent isoelectric points and relative percentages were determined. The relative CDT values (percent of total transferrin) matched expected values for both healthy and alcoholic samples. Because the method leveraged the sensitivity of an immunoassay, CDT was measured when serum samples were diluted up to 1200-fold, reducing the volume of serum required. Finally, the process is fully automated, with up to 96 samples analyzed per batch. |
| Databáze: | OpenAIRE |
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