Abstract
A significant population of ultrashort (50-150n) single-stranded DNA fragments were found in exosome-free blood plasma of retinoblastoma patients (6.84 ng mL-1), but not in plasma of healthy donors. An original high resolution HPLC technique has been proposed to reveal and characterize this peculiarity. To solve this task, a novel molecular size exclusion - anion exchange analytical technique was developed. Its applicability to diagnostics and oncogenesis research is quizzed here.
Highlights
The DNA repair machinery damage involves a marked DNA polymerase β hyperexpression and its processivity limitations in retinoblastoma cells (1, 2)
The content of ssDNA in the blood plasma of patients is 6.84 ± 0.56 ng mL−1, this DNA population consists of ultrashort fragments (50–150n)
An original procedure of size exclusion/anion exchange (SEAE)-HPLC we proposed can be used in biochemistry as well as in molecular biology, biotechnology, experimental oncology, analytical and clinical chemistry
Summary
The DNA repair machinery damage involves a marked DNA polymerase β hyperexpression and its processivity limitations in retinoblastoma cells (1, 2). It is hardly possible to overestimate this requirement since the conventional agarose gel electrophoresis and PCR procedures are not selective and sensitive enough as long as the pg-ng concentration range for short (smaller than 100n) ssDNA detection in clinical samples is the case (1–5). Taking this into account, the aim of a present work was to develop a new chromatographic technique suitable to solve the above specified task, a high resolution ssDNA analysis in liquid biopsies
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