Controlled proteolysis of normal and pathological prion protein in a microfluidic chip. - Institut Curie Accéder directement au contenu
Article Dans Une Revue Lab on a Chip Année : 2008

Controlled proteolysis of normal and pathological prion protein in a microfluidic chip.

Résumé

A microreactor for proteinase K (PK)-mediated protein digestion was developed as a step towards the elaboration of a fully integrated microdevice for the detection of pathological prion protein (PrP). PK-grafted magnetic beads were immobilized inside a polydimethylsiloxane (PDMS) microchannel using a longitudinal magnetic field parallel to the flow direction and a magnetic field gradient, thereby forming a matrix for enzymatic digestion. This self-organization provided uniform pore sizes, a low flow resistance and a strong reaction efficiency due to a very thin diffusion layer. The microreactor's performance was first evaluated using a model substrate, succinyl-ala-ala-ala-paranitroanilide (SAAAP). Reaction kinetics were typically accelerated a hundred-fold as compared to conventional batch reactions. Reproducibility was around 98% for on-chip experiments. This microsystem was then applied to the digestion of prion protein from brain tissues. Controlled proteolysis could be obtained by varying the on-chip flow rate, while a complete proteolysis of normal protein was achieved in only three minutes. Extracts from normal and pathological brain homogenates were finally compared and strong discrimination between normal and pathological samples was demonstrated.

Dates et versions

hal-00258875 , version 1 (25-02-2008)

Identifiants

Citer

Anne Le Nel, Nicolas Minc, Claire Smadja, Marcela Slovakova, Zuzana Bilkova, et al.. Controlled proteolysis of normal and pathological prion protein in a microfluidic chip.. Lab on a Chip, 2008, 8 (2), pp.294-301. ⟨10.1039/b715238h⟩. ⟨hal-00258875⟩
174 Consultations
0 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More