Cleavage Reaction Lateral Flow Assays for Salivary Pepsin Measurement Using a Pepsin-Susceptible Peptide Substrate

In this study, we introduce a novel cleavage reaction lateral flow assay (LFA) based on pepsin activity against a pepsin-susceptible peptide (PSP) substrate to detect salivary pepsin. Two types of cleavage reaction LFAs, the within-tube and on-strip cleavage reactions, were prepared based on the PSP...

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Main Authors: Sung-Woong Kang, Young Ju Lee, Jae-Chul Lee, Young-Gyu Eun, Gi-Ja Lee
Format: Article
Language:English
Published: MDPI AG 2024-11-01
Series:Chemosensors
Subjects:
Online Access:https://www.mdpi.com/2227-9040/12/11/241
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author Sung-Woong Kang
Young Ju Lee
Jae-Chul Lee
Young-Gyu Eun
Gi-Ja Lee
author_facet Sung-Woong Kang
Young Ju Lee
Jae-Chul Lee
Young-Gyu Eun
Gi-Ja Lee
author_sort Sung-Woong Kang
collection DOAJ
description In this study, we introduce a novel cleavage reaction lateral flow assay (LFA) based on pepsin activity against a pepsin-susceptible peptide (PSP) substrate to detect salivary pepsin. Two types of cleavage reaction LFAs, the within-tube and on-strip cleavage reactions, were prepared based on the PSP and pepsin reaction location. In the within-tube cleavage reaction LFA, samples were treated in the microtube within a heating block for 30 min separately and subsequently developed with running buffer in the LFA. For the on-strip cleavage reaction, samples were treated on the reaction zone of the strip within the heating zone of the multifunctional strip cassette for 10 min. After developing the running buffer in the LFA, the assay image was obtained using a universal mobile reader with a multifunctional strip cassette. The within-tube cleavage reaction LFA showed high sensitivity (limit of detection [LOD] 1.9 ng/mL), good specificity, and high reproducibility. This assay exhibited better linearity in the log concentration range of pepsin (4–500 ng/mL) than a commercially available dipstick assay. The on-strip cleavage reaction LFA showed a similar sensitivity (LOD 1.4 ng/mL) to that of the within-tube reaction assay. Therefore, we expect these cleavage reaction LFAs using PSP to be utilized as simple and effective tools to detect salivary pepsin.
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spelling doaj-art-fe2925877fb847efa66e2eb5b3e850d52024-11-26T17:57:15ZengMDPI AGChemosensors2227-90402024-11-01121124110.3390/chemosensors12110241Cleavage Reaction Lateral Flow Assays for Salivary Pepsin Measurement Using a Pepsin-Susceptible Peptide SubstrateSung-Woong Kang0Young Ju Lee1Jae-Chul Lee2Young-Gyu Eun3Gi-Ja Lee4Department of Medical Engineering, Graduate School, Kyung Hee University, Seoul 02447, Republic of KoreaDepartment of Biomedical Engineering, College of Medicine, Kyung Hee University, Seoul 02447, Republic of KoreaTextile Innovation R&D Department, Korea Institute of Industrial Technology, Ansan 15588, Republic of KoreaDepartment of Otolaryngology–Head and Neck Surgery, Kyung Hee University College of Medicine, Kyung Hee University Medical Center, Seoul 02447, Republic of KoreaDepartment of Medical Engineering, Graduate School, Kyung Hee University, Seoul 02447, Republic of KoreaIn this study, we introduce a novel cleavage reaction lateral flow assay (LFA) based on pepsin activity against a pepsin-susceptible peptide (PSP) substrate to detect salivary pepsin. Two types of cleavage reaction LFAs, the within-tube and on-strip cleavage reactions, were prepared based on the PSP and pepsin reaction location. In the within-tube cleavage reaction LFA, samples were treated in the microtube within a heating block for 30 min separately and subsequently developed with running buffer in the LFA. For the on-strip cleavage reaction, samples were treated on the reaction zone of the strip within the heating zone of the multifunctional strip cassette for 10 min. After developing the running buffer in the LFA, the assay image was obtained using a universal mobile reader with a multifunctional strip cassette. The within-tube cleavage reaction LFA showed high sensitivity (limit of detection [LOD] 1.9 ng/mL), good specificity, and high reproducibility. This assay exhibited better linearity in the log concentration range of pepsin (4–500 ng/mL) than a commercially available dipstick assay. The on-strip cleavage reaction LFA showed a similar sensitivity (LOD 1.4 ng/mL) to that of the within-tube reaction assay. Therefore, we expect these cleavage reaction LFAs using PSP to be utilized as simple and effective tools to detect salivary pepsin.https://www.mdpi.com/2227-9040/12/11/241pepsinsalivalateral flow assaycleavage reactionpepsin-susceptible peptide substratemultifunctional strip cassette
spellingShingle Sung-Woong Kang
Young Ju Lee
Jae-Chul Lee
Young-Gyu Eun
Gi-Ja Lee
Cleavage Reaction Lateral Flow Assays for Salivary Pepsin Measurement Using a Pepsin-Susceptible Peptide Substrate
Chemosensors
pepsin
saliva
lateral flow assay
cleavage reaction
pepsin-susceptible peptide substrate
multifunctional strip cassette
title Cleavage Reaction Lateral Flow Assays for Salivary Pepsin Measurement Using a Pepsin-Susceptible Peptide Substrate
title_full Cleavage Reaction Lateral Flow Assays for Salivary Pepsin Measurement Using a Pepsin-Susceptible Peptide Substrate
title_fullStr Cleavage Reaction Lateral Flow Assays for Salivary Pepsin Measurement Using a Pepsin-Susceptible Peptide Substrate
title_full_unstemmed Cleavage Reaction Lateral Flow Assays for Salivary Pepsin Measurement Using a Pepsin-Susceptible Peptide Substrate
title_short Cleavage Reaction Lateral Flow Assays for Salivary Pepsin Measurement Using a Pepsin-Susceptible Peptide Substrate
title_sort cleavage reaction lateral flow assays for salivary pepsin measurement using a pepsin susceptible peptide substrate
topic pepsin
saliva
lateral flow assay
cleavage reaction
pepsin-susceptible peptide substrate
multifunctional strip cassette
url https://www.mdpi.com/2227-9040/12/11/241
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