Related Experiment Video
Updated: Aug 14, 2026
09:56
Detection of Protease Activity by Fluorescent Peptide Zymography
Published on: January 20, 2019
Methodological considerations in GCF sampling with paper strips: poor recovery of uncomplexed elastase
1Department of Periodontology, Karolinska Institutet, Stockholm, Sweden.
Journal of Clinical Periodontology
|May 1, 1996
Summary
This study found most proteins are recovered well from paper strips for gingival crevicular fluid (GCF) sampling. However, free neutrophil elastase recovery was poor, impacting GCF analysis accuracy.
Area of Science:
- Biochemistry
- Periodontology
- Analytical Chemistry
Background:
- Gingival crevicular fluid (GCF) is a valuable source of biomarkers for periodontal disease.
- Accurate protein quantification from GCF samples is crucial for diagnosis and monitoring.
- Paper strips are commonly used for GCF sampling, but protein recovery efficiency needs evaluation.
Purpose of the Study:
- To assess the recovery rate of various proteins from paper strips used for GCF sampling.
- To evaluate the impact of elution buffer composition on protein recovery.
- To compare the performance of different paper strip materials for protein analysis.
Main Methods:
- Proteins (12-725 kD) were applied to paper strips and eluted using isotonic phosphate-buffered saline (pH 7.4) without detergent.
- Neutrophil lysis was avoided by using a non-lytic buffer.
- Recovery of free neutrophil elastase and elastase complexed with alpha-2-macroglobulin was specifically assessed.
- Polyvinylidene difluoride (Durapore) strips were tested as an alternative material.
- Measurements were compared using a Periotron 6000 device.
Main Results:
- Most proteins (approx. 90%) were satisfactorily recovered within 60 minutes.
- Elution buffer without detergent did not lyse neutrophils, preventing intracellular substance contamination.
- Free neutrophil elastase showed poor recovery, unlike elastase complexed with alpha-2-macroglobulin.
- Durapore strips yielded only 31% recovery of pure elastase and resulted in readings too low for accurate fluid volume measurements.
Conclusions:
- Isotonic buffers at neutral pH without detergents enable satisfactory and reproducible recovery of most proteins from paper strips.
- Protein recovery is generally independent of molecular weight within the tested range (12-725 kD).
- The poor recovery of uncomplexed neutrophil elastase is a significant limitation for its analysis in GCF samples collected on paper strips.
