NDLI: Rapid, continuous purification of proteins in a microfluidic device using genetically-engineered partition tags

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Front cover

Inside front cover

Contents and Chemical Technology

Research Highlights ( Lab on a Chip Vol - 8 Issue - 4 )

Glass coating for PDMS microfluidic channels by sol–gel methods

Digital microfluidics for cell-based assays

Rapid, continuous purification of proteins in a microfluidic device using genetically-engineered partition tags

Artificial cilia for active micro-fluidic mixing

A polymer lab-on-a-chip for magnetic immunoassay with on-chip sampling and detection capabilities

Parasitic trap cancellation using multiple frequency dielectrophoresis, demonstrated by loading cells into cages

Experimental and model investigation of the time-dependent 2-dimensional distribution of binding in a herringbone microchannel

Model based design of a microfluidic mixer driven by induced charge electroosmosis

Electrokinetics induced asymmetric transport in polymeric nanonozzles

Nano-scale superhydrophobicity: suppression of protein adsorption and promotion of flow-induced detachment

Rapid screening of membrane protein activity: electrophysiological analysis of OmpF reconstituted in proteoliposomes

Multiplexed proteomic sample preconcentration device using surface-patterned ion-selective membrane

Single molecule measurements within individual membrane-bound ion channels using a polymer-based bilayer lipid membrane chip

A low-cost, manufacturable method for fabricating capillary and optical fiber interconnects for microfluidic devices

Effect of the gelation process on the production of alginate microbeads by microfluidic chip technology

Shrinky-Dink microfluidics: 3D polystyrene chips

Non-equilibrium electrokinetic micro/nano fluidic mixer

Back matter

Back cover

Lab on a Chip Vol - 8 Issue - 3

Lab on a Chip Vol - 8 Issue - 2

Rapid, continuous purification of proteins in a microfluidic device using genetically-engineered partition tags

Content Provider	Royal Society of Chemistry (RSC)
Author	Light, Yooli K. Singh, Anup K. Meagher, Robert J.
Copyright Year	2008
Abstract	High-throughput screening assays of native and recombinant proteins are increasingly crucial in life science research, including fields such as drug screening and enzyme engineering. These assays are typically highly parallel, and require minute amounts of purified protein per assay. To address this need, we have developed a rapid, automated microscale process for isolating specific proteins from sub-microlitre volumes of E. Colicell lysate. Recombinant proteins are genetically tagged to drive partitioning into the PEG-rich phase of a flowing aqueous two-phase system, which removes ∼85% of contaminating proteins, as well as unwanted nucleic acids and cell debris, on a simple microfluidic device. Inclusion of the genetic tag roughly triples recovery of the autofluorescent protein AcGFP1, and also significantly improves recovery of the enzyme glutathione S-transferase (GST), from nearly zero recovery for the wild-type enzyme, up to 40% with genetic tagging. The extraction process operates continuously, with only a single step from cell lysate to purified protein, and does not require expensive affinity reagents or troublesome chromatographic steps. The two-phase system is mild and does not disrupt protein function, as evidenced by recovery of active enzymes and functional fluorescent protein from our microfluidic process. The microfluidic aqueous two-phase extraction forms the core component of an integrated lab-on-a-chip device comprising cell culture, lysis, purification and analysis on a single device.
Starting Page	527
Ending Page	532
Page Count	6
File Format	HTM / HTML PDF
ISSN	14730197
Volume Number	8
Issue Number	4
Journal	Lab on a Chip
DOI	10.1039/b716462a
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Cell culture Enzyme Green fluorescent protein Lysis High-throughput screening Lab-on-a-chip Glutathione S-transferase Liquid\u2013liquid extraction Protein Aqueous two-phase system Chromatography
Content Type	Text
Resource Type	Article
Subject	Chemistry Nanoscience and Nanotechnology Biochemistry Bioengineering Biomedical Engineering

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Integrated microfluidic cell culture and lysis on a chip

Continuous dielectrophoretic cell separation microfluidic device

On-chip screening method for cell migration genes based on a transfection microarray

Self-assembly of glutathione S-transferase into nanowires

Observations of green fluorescent protein as a fusion partner in genetically engineered Escherichia coli: Monitoring protein expression and solubility

Lab-on-a-chip-based high-throughput screening of the genotoxicity of engineered nanomaterials.

ElectroTaxis-on-a-Chip (ETC): an integrated quantitative high-throughput screening platform for electrical field-directed cell migration

Examination of laser microbeam cell lysis in a PDMS microfluidic channel using time-resolved imaging

Rapid self-assembly of DNA on a microfluidic chip

Rapid, continuous purification of proteins in a microfluidic device using genetically-engineered partition tags

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