On-a-chip tryptic digestion of transthyretin: A step toward an integrated microfluidic system for the follow-up of familial transthyretin amyloidosis

Research output: Contribution to journalJournal articleResearchpeer-review

Standard

On-a-chip tryptic digestion of transthyretin : A step toward an integrated microfluidic system for the follow-up of familial transthyretin amyloidosis. / Bataille, Jeanne; Viodé, Arthur; Pereiro, Iago; Lafleur, Josiane P.; Varenne, Fanny; Descroix, Stéphanie; Becher, François; Kutter, Jörg P.; Roesch, Camille; Poüs, Christian; Taverna, Myriam; Pallandre, Antoine; Smadja, Claire; Le Potier, Isabelle.

In: Analyst, Vol. 143, No. 5, 07.03.2018, p. 1077-1086.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Bataille, J, Viodé, A, Pereiro, I, Lafleur, JP, Varenne, F, Descroix, S, Becher, F, Kutter, JP, Roesch, C, Poüs, C, Taverna, M, Pallandre, A, Smadja, C & Le Potier, I 2018, 'On-a-chip tryptic digestion of transthyretin: A step toward an integrated microfluidic system for the follow-up of familial transthyretin amyloidosis', Analyst, vol. 143, no. 5, pp. 1077-1086. https://doi.org/10.1039/c7an01737e

APA

Bataille, J., Viodé, A., Pereiro, I., Lafleur, J. P., Varenne, F., Descroix, S., Becher, F., Kutter, J. P., Roesch, C., Poüs, C., Taverna, M., Pallandre, A., Smadja, C., & Le Potier, I. (2018). On-a-chip tryptic digestion of transthyretin: A step toward an integrated microfluidic system for the follow-up of familial transthyretin amyloidosis. Analyst, 143(5), 1077-1086. https://doi.org/10.1039/c7an01737e

Vancouver

Bataille J, Viodé A, Pereiro I, Lafleur JP, Varenne F, Descroix S et al. On-a-chip tryptic digestion of transthyretin: A step toward an integrated microfluidic system for the follow-up of familial transthyretin amyloidosis. Analyst. 2018 Mar 7;143(5):1077-1086. https://doi.org/10.1039/c7an01737e

Author

Bataille, Jeanne ; Viodé, Arthur ; Pereiro, Iago ; Lafleur, Josiane P. ; Varenne, Fanny ; Descroix, Stéphanie ; Becher, François ; Kutter, Jörg P. ; Roesch, Camille ; Poüs, Christian ; Taverna, Myriam ; Pallandre, Antoine ; Smadja, Claire ; Le Potier, Isabelle. / On-a-chip tryptic digestion of transthyretin : A step toward an integrated microfluidic system for the follow-up of familial transthyretin amyloidosis. In: Analyst. 2018 ; Vol. 143, No. 5. pp. 1077-1086.

Bibtex

@article{9b186cf1039343e7bcf613f101edeabb,
title = "On-a-chip tryptic digestion of transthyretin: A step toward an integrated microfluidic system for the follow-up of familial transthyretin amyloidosis",
abstract = "A microfluidic microreactor for trypsin mediated transthyretin (TTR) digestion has been developed as a step towards the elaboration of a fully integrated microdevice for the detection of a rare and disabling disease, the familial transthyretin amyloidosis (ATTR) which is related to specific TTR mutations. Therefore, an enzymatic microreactor coupled to an analytical step able to monitor the mutation of TTR on specific peptide fragments would allow an accurate monitoring of the treatment efficiency of ATTR. In this study, two types of immobilized trypsin microreactors have been investigated: a new miniaturized, microfluidic fluidized bed packed with trypsin functionalized magnetic particles (MPs), and a thiol-ene (TE) monolith-based chip. Their performances were first demonstrated with N-benzoyl-dl-arginine-4-nitroanilide hydrochloride BApNA, a low molecular weight substrate. High reaction yields (75.2%) have been reached within 0.6 min for the TE-based trypsin microreactor, while a lower yield (12.4%) was obtained for the micro-fluidized bed within a similar residence time. Transposition of the optimized conditions, developed with BApNA, to TTR digestion in the TE-based trypsin microreactor was successfully performed. We demonstrated that the TE-chip can achieve an efficient and reproducible digestion of TTR. This has been assessed by MS detection. In addition, TTR hydrolysis led to the production of a fragment of interest allowing the therapeutic follow-up of more than twenty possible ATTR mutations. High sequence coverage (90%), similar to those obtained with free trypsin, was achieved in a short time (2.4 min). Repeated experiments showed good reproducibility (RSD = 6.8%). These promising results open up the route for an innovative treatment follow-up dedicated to ATTR.",
author = "Jeanne Bataille and Arthur Viod{\'e} and Iago Pereiro and Lafleur, {Josiane P.} and Fanny Varenne and St{\'e}phanie Descroix and Fran{\c c}ois Becher and Kutter, {J{\"o}rg P.} and Camille Roesch and Christian Po{\"u}s and Myriam Taverna and Antoine Pallandre and Claire Smadja and {Le Potier}, Isabelle",
year = "2018",
month = mar,
day = "7",
doi = "10.1039/c7an01737e",
language = "English",
volume = "143",
pages = "1077--1086",
journal = "The Analyst",
issn = "0003-2654",
publisher = "Royal Society of Chemistry",
number = "5",

}

RIS

TY - JOUR

T1 - On-a-chip tryptic digestion of transthyretin

T2 - A step toward an integrated microfluidic system for the follow-up of familial transthyretin amyloidosis

AU - Bataille, Jeanne

AU - Viodé, Arthur

AU - Pereiro, Iago

AU - Lafleur, Josiane P.

AU - Varenne, Fanny

AU - Descroix, Stéphanie

AU - Becher, François

AU - Kutter, Jörg P.

AU - Roesch, Camille

AU - Poüs, Christian

AU - Taverna, Myriam

AU - Pallandre, Antoine

AU - Smadja, Claire

AU - Le Potier, Isabelle

PY - 2018/3/7

Y1 - 2018/3/7

N2 - A microfluidic microreactor for trypsin mediated transthyretin (TTR) digestion has been developed as a step towards the elaboration of a fully integrated microdevice for the detection of a rare and disabling disease, the familial transthyretin amyloidosis (ATTR) which is related to specific TTR mutations. Therefore, an enzymatic microreactor coupled to an analytical step able to monitor the mutation of TTR on specific peptide fragments would allow an accurate monitoring of the treatment efficiency of ATTR. In this study, two types of immobilized trypsin microreactors have been investigated: a new miniaturized, microfluidic fluidized bed packed with trypsin functionalized magnetic particles (MPs), and a thiol-ene (TE) monolith-based chip. Their performances were first demonstrated with N-benzoyl-dl-arginine-4-nitroanilide hydrochloride BApNA, a low molecular weight substrate. High reaction yields (75.2%) have been reached within 0.6 min for the TE-based trypsin microreactor, while a lower yield (12.4%) was obtained for the micro-fluidized bed within a similar residence time. Transposition of the optimized conditions, developed with BApNA, to TTR digestion in the TE-based trypsin microreactor was successfully performed. We demonstrated that the TE-chip can achieve an efficient and reproducible digestion of TTR. This has been assessed by MS detection. In addition, TTR hydrolysis led to the production of a fragment of interest allowing the therapeutic follow-up of more than twenty possible ATTR mutations. High sequence coverage (90%), similar to those obtained with free trypsin, was achieved in a short time (2.4 min). Repeated experiments showed good reproducibility (RSD = 6.8%). These promising results open up the route for an innovative treatment follow-up dedicated to ATTR.

AB - A microfluidic microreactor for trypsin mediated transthyretin (TTR) digestion has been developed as a step towards the elaboration of a fully integrated microdevice for the detection of a rare and disabling disease, the familial transthyretin amyloidosis (ATTR) which is related to specific TTR mutations. Therefore, an enzymatic microreactor coupled to an analytical step able to monitor the mutation of TTR on specific peptide fragments would allow an accurate monitoring of the treatment efficiency of ATTR. In this study, two types of immobilized trypsin microreactors have been investigated: a new miniaturized, microfluidic fluidized bed packed with trypsin functionalized magnetic particles (MPs), and a thiol-ene (TE) monolith-based chip. Their performances were first demonstrated with N-benzoyl-dl-arginine-4-nitroanilide hydrochloride BApNA, a low molecular weight substrate. High reaction yields (75.2%) have been reached within 0.6 min for the TE-based trypsin microreactor, while a lower yield (12.4%) was obtained for the micro-fluidized bed within a similar residence time. Transposition of the optimized conditions, developed with BApNA, to TTR digestion in the TE-based trypsin microreactor was successfully performed. We demonstrated that the TE-chip can achieve an efficient and reproducible digestion of TTR. This has been assessed by MS detection. In addition, TTR hydrolysis led to the production of a fragment of interest allowing the therapeutic follow-up of more than twenty possible ATTR mutations. High sequence coverage (90%), similar to those obtained with free trypsin, was achieved in a short time (2.4 min). Repeated experiments showed good reproducibility (RSD = 6.8%). These promising results open up the route for an innovative treatment follow-up dedicated to ATTR.

UR - http://www.scopus.com/inward/record.url?scp=85042698351&partnerID=8YFLogxK

U2 - 10.1039/c7an01737e

DO - 10.1039/c7an01737e

M3 - Journal article

C2 - 29383369

AN - SCOPUS:85042698351

VL - 143

SP - 1077

EP - 1086

JO - The Analyst

JF - The Analyst

SN - 0003-2654

IS - 5

ER -

ID: 223823749