Detection of β-lactoglobulin under different thermal-processing conditions by immunoassay based on nanobody and monoclonal antibody

Shijie Li; Linqing Nie; Yi Wang; YaYa Wang; Dancai Fan; Junping Wang; Yaozhong Hu; Lu Dong; Yan Zhang; Shuo Wang

doi:10.1016/j.foodchem.2023.136337

Detection of β-lactoglobulin under different thermal-processing conditions by immunoassay based on nanobody and monoclonal antibody

Food Chem. 2023 Oct 30:424:136337. doi: 10.1016/j.foodchem.2023.136337. Epub 2023 May 9.

Authors

Shijie Li¹, Linqing Nie², Yi Wang³, YaYa Wang¹, Dancai Fan¹, Junping Wang⁴, Yaozhong Hu⁵, Lu Dong⁶, Yan Zhang⁷, Shuo Wang⁸

Affiliations

¹ Tianjin Key Laboratory of Food Science and Health, School of Medicine, Nankai University, Tianjin 300071, China.
² Tianjin Key Laboratory of Food Science and Health, School of Medicine, Nankai University, Tianjin 300071, China. Electronic address: 120211403@mail.nankai.edu.cn.
³ Tianjin Key Laboratory of Food Science and Health, School of Medicine, Nankai University, Tianjin 300071, China. Electronic address: 2120201268@mail.nankai.edu.cn.
⁴ State Key Laboratory of Food Nutrition and Safety, Key Laboratory of Food Nutrition and Safety, Ministry of Education of China, Tianjin University of Science and Technology, Tianjin 300457, China. Electronic address: wangjp@tust.edu.cn.
⁵ Tianjin Key Laboratory of Food Science and Health, School of Medicine, Nankai University, Tianjin 300071, China. Electronic address: yzhu@nankai.edu.cn.
⁶ Tianjin Key Laboratory of Food Science and Health, School of Medicine, Nankai University, Tianjin 300071, China. Electronic address: 819063@nankai.edu.cn.
⁷ Tianjin Key Laboratory of Food Science and Health, School of Medicine, Nankai University, Tianjin 300071, China. Electronic address: yzhang@nankai.edu.cn.
⁸ Tianjin Key Laboratory of Food Science and Health, School of Medicine, Nankai University, Tianjin 300071, China. Electronic address: wangshuo@nankai.edu.cn.

PMID: 37209435
DOI: 10.1016/j.foodchem.2023.136337

Abstract

The problems of inaccurate detection values of thermal-processed β-lactoglobulin (β-LG) content seriously affect the screening of allergens. A monoclonal antibody (mAb) against β-LG was successfully prepared and a highly sensitive sandwich ELISA (sELISA) was constructed with specific nanobody (Nb) as the capture antibody with detection limit of 0.24 ng/mL. Based on this sELISA, the ability of Nb and mAb to recognize β-LG and β-LG interacting with milk components was explored. Combined with protein structure analysis to elaborate the mechanism of shielding β-LG antigen epitopes during thermal-processing, thus enabling the differentiation between pasteurized and ultra-high temperature sterilized milk, the detection of milk content in milk-containing beverages, and the highly sensitive detection and analysis of β-LG allergens in dairy-free products. The method provides methodological support for identifying the quality of dairy products and reducing the risk of β-LG contamination in dairy-free products.

Keywords: Allergens detection; Discrimination mechanism analysis; Nanobody; Processed milk; β-lactoglobulin.

MeSH terms

Allergens / analysis
Animals
Antibodies, Monoclonal*
Enzyme-Linked Immunosorbent Assay / methods
Immunoassay
Lactoglobulins* / chemistry
Milk / chemistry

Substances

Antibodies, Monoclonal
Lactoglobulins
Allergens