A comparison of two commercially available ELISA methods for the quantification of human plasma heat shock protein 70 during rest and exercise stress
- Published
- Accepted
- Subject Areas
- Biochemistry, Anatomy and Physiology
- Keywords
- Heat shock proteins (HSP), acute heat stress, human, acute hypoxia, ELISA comparision, exercise
- Copyright
- © 2015 Lee et al.
- Licence
- This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ PrePrints) and either DOI or URL of the article must be cited.
- Cite this article
- 2015. A comparison of two commercially available ELISA methods for the quantification of human plasma heat shock protein 70 during rest and exercise stress. PeerJ PrePrints 3:e1120v1 https://doi.org/10.7287/peerj.preprints.1120v1
Abstract
Background. This study compared resting and exercise heat/hypoxic-stress induced levels of plasma eHSP70 in humans using two commercially available ELISA kits. Methods. EDTA plasma samples were collected from 21 males during two separate investigations. Participants in Part A completed a 60 min treadmill run in the heat (HOT70; 33.0 ± 0.1 °C, 28.7 ± 0.8%, n = 6) at 70% V̇O2max. Participants in Part B completed 60 minutes of cycling exercise at 50% V̇O2max in either hot (HOT50; 40.5°C, 25.4 RH%, n = 7) or hypoxic (HYP50; FIO2 = 0.14, 21°C, 35% RH, n = 8) conditions. Samples were collected prior to and immediately upon termination of exercise and analysed for eHSP70 using EKS-715 high sensitivity HSP70 ELISA, and new ENZ-KIT-101 AMP’D™ HSP70 high sensitivity ELISA. Results. ENZ-KIT was superior in detecting resting eHSP70 (1.54 ± 3.27 ng.mL-1; range 0.08 to 14.01 ng.mL-1), with concentrations obtained from 100% of samples compared to 19% with EKS-715 assay. The ENZ-KIT requires optimisation prior to running samples in order to ensure participants fall within the standard curve, a step not required with EKS-715. Using ENZ-KIT, a 1:4 dilution allowed for quantification of resting HSP70 in 26/32 samples, with a 1:8 (n = 3) and 1:16 (n = 3) dilution required to determine the remaining samples. After exercise eHSP70 was detected in 6/21 and 21/21 samples using EKS-715 and ENZ-KIT respectively. eHSP70 was increased from rest after HOT70 (p < 0.05), but not HOT50 (p > 0.05) or HYP50 (p > 0.05) when analysed using ENZ-KIT. Conclusion. It is recommended that future studies requiring the precise determination of resting plasma eHSP70 use the ENZ-KIT (i.e., HSP70 Amp'd®ELISA) instead of the EKS-715 assay, despite additional assay development time and cost required.
Author Comment
This paper is the first version of a paper submitted Cell Stress and Chaperones. The data presented herein provides important data for researchers measuring circulating HSP70 in humans.