Does Transglutaminase Crosslinking Reduce the Antibody Recognition Capacity of β-Lactoglobulin: An Analysis from Conformational Perspective

Food allergies are a global concern, with β-lactoglobulin (β-LG) in bovine milk being a major allergenic protein. This study investigated the effects of transglutaminase (TGase)-mediated crosslinking on the antibody recognition capacity (ARC) and structural properties of β-LG, with the aim of develo...

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Main Authors: Lei Fang, Xun Han, Yue Zhang, Tianran Hui, Lingling Ding, Wenlu Dai, Yujie Han, Maoqiang Zheng, Guangliang Xing
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Molecules
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Online Access:https://www.mdpi.com/1420-3049/30/3/685
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author Lei Fang
Xun Han
Yue Zhang
Tianran Hui
Lingling Ding
Wenlu Dai
Yujie Han
Maoqiang Zheng
Guangliang Xing
author_facet Lei Fang
Xun Han
Yue Zhang
Tianran Hui
Lingling Ding
Wenlu Dai
Yujie Han
Maoqiang Zheng
Guangliang Xing
author_sort Lei Fang
collection DOAJ
description Food allergies are a global concern, with β-lactoglobulin (β-LG) in bovine milk being a major allergenic protein. This study investigated the effects of transglutaminase (TGase)-mediated crosslinking on the antibody recognition capacity (ARC) and structural properties of β-LG, with the aim of developing hypoallergenic dairy products. β-LG solutions were treated with TGase at varying concentrations (0, 5, 10, 15, and 20 U/g) and durations (0, 6, 18, 24, and 42 h), followed by analysis using electrophoresis, enzyme-linked immunosorbent assay (ELISA), and spectroscopy. The results demonstrated that treatment with TGase at 20 U/g significantly reduced the ARC and immunoglobulin E (IgE) binding capacity of β-LG to 90.0 ± 0.4% and 58.4 ± 1.0%, respectively, with the optimal ARC reduction observed after 6 h of treatment (86.7 ± 1.2%, <i>p</i> < 0.05). Although electrophoresis did not reveal significant crosslinking of β-LG, ultraviolet absorption, fluorescence intensity, and hydrophobicity all increased with prolonged crosslinking time, while sulfhydryl content fluctuated irregularly. These findings suggest that β-LG underwent varying degrees of structural modification, which led to the masking of antigenic epitopes during the early stages (0–24 h), followed by their re-exposure at the later stage (42 h). Overall, these results highlight the potential of TGase to reduce β-LG potential allergenicity, presenting a promising strategy for the development of hypoallergenic dairy products.
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spelling doaj-art-01c0d69d1400401ba7988091d39a3d6d2025-08-20T02:12:31ZengMDPI AGMolecules1420-30492025-02-0130368510.3390/molecules30030685Does Transglutaminase Crosslinking Reduce the Antibody Recognition Capacity of β-Lactoglobulin: An Analysis from Conformational PerspectiveLei Fang0Xun Han1Yue Zhang2Tianran Hui3Lingling Ding4Wenlu Dai5Yujie Han6Maoqiang Zheng7Guangliang Xing8School of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, ChinaSchool of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, ChinaSchool of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, ChinaUCL Division of Medicine, University College London, London WC1E 6BT, UKSchool of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, ChinaSchool of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, ChinaSchool of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, ChinaSchool of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, ChinaSchool of Biology and Food Engineering, Changshu Institute of Technology, Changshu 215500, ChinaFood allergies are a global concern, with β-lactoglobulin (β-LG) in bovine milk being a major allergenic protein. This study investigated the effects of transglutaminase (TGase)-mediated crosslinking on the antibody recognition capacity (ARC) and structural properties of β-LG, with the aim of developing hypoallergenic dairy products. β-LG solutions were treated with TGase at varying concentrations (0, 5, 10, 15, and 20 U/g) and durations (0, 6, 18, 24, and 42 h), followed by analysis using electrophoresis, enzyme-linked immunosorbent assay (ELISA), and spectroscopy. The results demonstrated that treatment with TGase at 20 U/g significantly reduced the ARC and immunoglobulin E (IgE) binding capacity of β-LG to 90.0 ± 0.4% and 58.4 ± 1.0%, respectively, with the optimal ARC reduction observed after 6 h of treatment (86.7 ± 1.2%, <i>p</i> < 0.05). Although electrophoresis did not reveal significant crosslinking of β-LG, ultraviolet absorption, fluorescence intensity, and hydrophobicity all increased with prolonged crosslinking time, while sulfhydryl content fluctuated irregularly. These findings suggest that β-LG underwent varying degrees of structural modification, which led to the masking of antigenic epitopes during the early stages (0–24 h), followed by their re-exposure at the later stage (42 h). Overall, these results highlight the potential of TGase to reduce β-LG potential allergenicity, presenting a promising strategy for the development of hypoallergenic dairy products.https://www.mdpi.com/1420-3049/30/3/685β-lactoglobulintransglutaminaseantibody recognitionconformational changes
spellingShingle Lei Fang
Xun Han
Yue Zhang
Tianran Hui
Lingling Ding
Wenlu Dai
Yujie Han
Maoqiang Zheng
Guangliang Xing
Does Transglutaminase Crosslinking Reduce the Antibody Recognition Capacity of β-Lactoglobulin: An Analysis from Conformational Perspective
Molecules
β-lactoglobulin
transglutaminase
antibody recognition
conformational changes
title Does Transglutaminase Crosslinking Reduce the Antibody Recognition Capacity of β-Lactoglobulin: An Analysis from Conformational Perspective
title_full Does Transglutaminase Crosslinking Reduce the Antibody Recognition Capacity of β-Lactoglobulin: An Analysis from Conformational Perspective
title_fullStr Does Transglutaminase Crosslinking Reduce the Antibody Recognition Capacity of β-Lactoglobulin: An Analysis from Conformational Perspective
title_full_unstemmed Does Transglutaminase Crosslinking Reduce the Antibody Recognition Capacity of β-Lactoglobulin: An Analysis from Conformational Perspective
title_short Does Transglutaminase Crosslinking Reduce the Antibody Recognition Capacity of β-Lactoglobulin: An Analysis from Conformational Perspective
title_sort does transglutaminase crosslinking reduce the antibody recognition capacity of β lactoglobulin an analysis from conformational perspective
topic β-lactoglobulin
transglutaminase
antibody recognition
conformational changes
url https://www.mdpi.com/1420-3049/30/3/685
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