Development Of Neatstik - A Novel Lateral Flow Device For The Detection Of Active Neutrophil Elastase In Patients With Respiratory Disease

Kelly Moffitt, W Tong, Timothy Ferguson, Darragh McCafferty, Charlene Robb, Lorraine Martin, Brian Walker

Research output: Contribution to journalMeeting abstractpeer-review

Abstract

Introduction: Chronic respiratory diseases such as COPD, bronchiectasis and cystic fibrosis (CF), are a huge burden on a significant proportion of the population. It's a well-accepted concept within the scientific and medical community that active neutrophil elastase (NE) represents a strong biomarker for monitoring the severity and progression of a number of chronic respiratory diseases. Using our novel ProteaseTag® technology, we have already developed a laboratory-use Activity-Based Immunoassay (ABI) for the detection and measurement of active NE in clinical samples. However, rapid measurement of NE activity at the point-of-care (such as in the clinic) would provide important information, which alongside lung function data, could help to determine the extent of exacerbation and assist in the prescribing of antibiotics. Therefore, we are in the process of converting our NE-ABI technology into a lateral flow, point-of-care (PoC) test for measuring active NE, which would provide a result in under 15 minutes, and would be suitable for use within the clinic. We envisage that NEATstik (Neutrophil Elastase Airways Test) will allow for routine monitoring of patient samples, improve patient outcome and reduce the clinical, societal and economic burdens associated with chronic respiratory disease.
Methods: The concentration of active NE in CF sputum sol was determined using the ProteaseTag® Active NE Activity Based Immunoassay (ProAxsis Ltd). This allowed the identification of samples containing active NE ranging from 2.75ng/ml-4400 ng/ml, which were then selected for measurement by the PoC prototype. Each test strip was dropped into a glass tube containing 75 μl of sputum sol diluted in running buffer. The sample was left to run for 15 minutes and a visual grade of the resulting test line scored between 0-10.
Results: Test lines were visible for all samples containing NE. There was no ‘non-specific binding’ on the strips tested with buffer only. In this format, the test grade lines were proportionate in intensity at NE concentrations of 2.75 ng/ml (visual grade 2) – 4400 ng/ml (visual grade 8). This detection range can be modified as appropriate by changing the concentration of the reagents and/or the components of the device.
Conclusion: The development of the first highly sensitive and specific lateral flow device for the rapid detection of active NE in complex clinical samples will enable proactive management of multiple chronic respiratory diseases. In particular, it has the potential to assist in the prediction of upcoming exacerbations, and thus allow pre-emptive treatment to minimise their impact.
Original languageEnglish
Article numberA6309
Number of pages1
JournalAmerican Journal of Respiratory and Critical Care Medicine
Volume195
Publication statusPublished - 22 May 2017
EventAmerican Thoracic Society (ATS) 2017: ATS 2017 - Washington DC, United States
Duration: 21 May 201724 Jan 2018

Keywords

  • neutrophil elastase
  • point-of-care test
  • CF
  • COPD

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