Enhanced microdialysis relative recovery of inflammatory cytokines using antibody-coated microspheres analyzed by flow cytometry

Analytical Chemistry
Xiaoping AoJ A Stenken

Abstract

Achieving high relative recovery (RR) of proteins during microdialysis sampling is difficult due to the diffusion limitations inherent to this sampling process. This often causes low microdialysis RR for proteins with molecular weight >10 kDa. A RR enhancement process for microdialysis sampling of proteins has been developed that can be readily used with flow cytometers. Multiplexed RR enhancement and detection of five different cytokines (TNF-alpha, IFN-gamma, IL-2, IL-4, and IL-5) was achieved by including antibody-coated microspheres in the microdialysis perfusion fluid. Inclusion of these antibody-coated microspheres causes an increase in the analyte diffusive driving force across the dialysis membrane and a subsequent increase in the relative recovery. For the five cytokines, typical control and enhanced relative recoveries at a 1.0 microL/min flow rate were as follows (n = 3): TNF-alpha, 5.5 +/- 0.6% and 60.4 +/- 5.8%; IFN-gamma, 2.6 +/- 0.3% and 25.8 +/- 2.3%; IL-5, 1.4 +/- 0.2% and 4.9 +/- 0.1%; IL-4, 10.9 +/- 0.6% and 78.8 +/- 8.0%; and IL-2, 4.1 +/- 0.4% and 19.8 +/- 2.5%. Using this approach, a four- to 12-fold enhancement of microdialysis RR was achieved for the five cytokines from a quiescent solution. The enhancem...Continue Reading

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Citations

Nov 6, 2010·Analytical and Bioanalytical Chemistry·Jia Duo, Julie A Stenken
Jun 7, 2005·Journal of Microbiological Methods·Xiaoping AoJulie A Stenken
Dec 16, 2005·The AAPS Journal·Geraldine F Clough
Jun 19, 2013·Annual Review of Analytical Chemistry·Michelle L Rogers, Martyn G Boutelle
Apr 22, 2014·Journal of Pharmaceutical and Biomedical Analysis·Lin LiuXingguo Zhang
Jan 18, 2006·Journal of Immunological Methods·A J RosenbloomT L Whiteside
Nov 26, 2013·European Journal of Pharmaceutical Sciences : Official Journal of the European Federation for Pharmaceutical Sciences·Cynthia R Sides, Julie A Stenken
Sep 28, 2016·Journal of Pharmaceutical Sciences·Satyawan B JadhavHartmut Derendorf
Jul 13, 2019·Journal of Neurotrauma·Susan Giorgi-CollAdel Helmy
Apr 21, 2007·Clinical Chemistry·Yuan-Yuan LiShun-Qing Xu
Oct 16, 2004·Analytical Chemistry·Robert J SchutteW Monty Reichert

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