BIABooster: Online DNA Concentration and Size Profiling with a Limit of Detection of 10 fg/μL and Application to High-Sensitivity Characterization of Circulating Cell-Free DNA

Comtet-Louis Andriamanampisoa 1 Aurélien Bancaud 2 Audrey Boutonnet-Rodat 1 Audrey Didelot 3 Jacques Fabre 4 Frederic Fina 5 Fanny Garlan 6 Sonia Garrigou 6 Caroline Gaudy 7 Frédéric Ginot 1 Daniel Henaff 8 Pierre Laurent-Puig 9 Arnaud Morin 10 Vincent Picot 1 Laure Saias 11 Valérie Taly 12 Pascale Tomasini Aziz Zaanan 13
1 Picometrics Technologies
2 LAAS-MILE - Équipe Micro-Nanofluidique pour les sciences de la vie et de l’environnement
LAAS - Laboratoire d'analyse et d'architecture des systèmes
3 EPHE - École pratique des hautes études
4 Université d'Amiens
5 Laboratoire de Transfert d'Oncologie Biologique [Hôpital Nord - APHM]
6 MEPPOT - U1147 - Médecine Personnalisée, Pharmacogénomique, Optimisation Thérapeutique
7 LSIS - Laboratoire des Sciences de l'Information et des Systèmes
8 IGMM - Institut de Génétique Moléculaire de Montpellier
9 Unicancer
10 LCPEM - Laboratoire des Composants PEM
11 PCC - Physico-Chimie-Curie
12 CRC (UMR_S_1138 / U1138) - Centre de Recherche des Cordeliers
13 UPD5 - Université Paris Descartes - Paris 5
Abstract : We describe a technology to perform sizing and concentration analysis of double stranded DNA with a sensitivity of 10 fg/μL in an operating time of 20 min. The technology is operated automatically on a commercial capillary electrophoresis instrument using electro-hydrodynamic actuation. It relies on a new capillary device that achieves online concentration of DNA at the junction between two capillaries of different diameters, thanks to viscoelastic lift forces. Using a set of DNA ladders in the range of 100–1500 bp, we report a sizing accuracy and precision better than 3% and a concentration quantification precision of ∼20%. When the technology is applied to the analysis of clinical samples of circulating cell-free DNA (cfDNA), the measured cfDNA concentrations are in good correlation with those measured by digital PCR. Furthermore, the cfDNA size profiles indicate that the fraction of low molecular weight cfDNA in the range of 75–240 bp is a candidate biomarker to discriminate between healthy subjects and cancer patients. We conclude that our technology is efficient in analyzing highly diluted DNA samples and suggest that it will be helpful in translational and clinical research involving cfDNA.
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UNIV-TLSE3 | CURIE | LSIS | CNRS | CEA | FNCLCC | INC-CNRS | UNIV-PARIS5 | BS | UNIV-MONTPELLIER | INSA-TOULOUSE | LIS-LAB | UNIV-TLN | UNIV-AMU | INSA-GROUPE | SORBONNE-UNIVERSITE | UNIV-PARIS7 | PCC_UMR168 | CORDELIERS | EPHE | PSL | LAAS-MILE | LAAS | LAAS-SYSTEMES-COMPLEXES | USPC | UNIV-HESAM | SU-SCIENCES | UT1-CAPITOLE | UNIV-TLSE2

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Comtet-Louis Andriamanampisoa, Aurélien Bancaud, Audrey Boutonnet-Rodat, Audrey Didelot, Jacques Fabre, et al.. BIABooster: Online DNA Concentration and Size Profiling with a Limit of Detection of 10 fg/μL and Application to High-Sensitivity Characterization of Circulating Cell-Free DNA. Analytical Chemistry, American Chemical Society, 2018, 90 (6), pp.3766-3774. ⟨10.1021/acs.analchem.7b04034⟩. ⟨inserm-02304850⟩

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