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New multi-step kinetics using common affinity biosensors saves time and sample at full access to kinetics and concentration

Paper ID Volume ID Publish Year Pages File Format Full-Text
25447 43574 2006 5 PDF Available
Title
New multi-step kinetics using common affinity biosensors saves time and sample at full access to kinetics and concentration
Abstract

Today, affinity-based biosensorics is a standard technology in quantitative biomolecular interaction analysis, but suffers from low sample throughput and sometimes from inaccessible kinetics. A new methodology for such biosensors is introduced here that cuts down measurement time dramatically and increases confidentiality of results. In contrast to traditional applications, the ligand immobilized on the sensor chip is exposed to the binding analyte at a rapid stepwise change of the analyte concentration without the need for regenerations between analyte additions. In the application presented here, each addition of the analyte is succeeded by a buffer flow, yielding alternating association and dissociation phases in a “zigzag” style. This binding curve pattern is analyzed by means of novel fitting algorithms, which render detailed kinetics rate constants at a high level of self-consistency, and hence, validity due to multiple cross-checks. In comparison with traditional sequential kinetics analysis, this new multi-step kinetics approach returns practically identical (or improved) kinetics constants—at valuable savings in time/material since regeneration steps, ligand re-captures, or titration equilibrations are unnecessary.

Keywords
GST, glutathione S-transferase; GST-PKI, GST-fused heat-stable protein kinase inhibitor of cAMP-dependent protein kinase; MTL, mass-transport limitation(s); mNIAR, molar net-initial association rate; mNFAR, molar net-final association rate; MSK, multi-ste
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New multi-step kinetics using common affinity biosensors saves time and sample at full access to kinetics and concentration
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 124, Issue 1, 25 June 2006, Pages 191–195
Authors
,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us