Abstract
Non-specific lipid transfer protein (LTP) in barley grain reacted with the IgE in sera drawn from food allergy patients. A sandwich-type of enzyme-linked immunosorbent assay (ELISA) was developed with mouse monoclonal antibodies raised against LTP purified with barley flour. This ELISA showed a practical working range of 0.3–3 ng/mL and no cross-reactivity with wheat, adlay and rye. Using this ELISA, LTP was determined in several types of barley-foods, including fermented foods such as malt vinegar, barley-malt miso and beer. LTP content in beer of the same kind was approximately constant, even if manufacturing factory and production days were different. Not only as a factor of foam formation and stability but also as an allergen, controlling and monitoring of LTP in beer should be considered. Taken together, our LTP-detecting ELISA can be proposed as an appropriate system for the quality control of beer.
Highlights
Non-specific lipid transfer protein (LTP) is an ubiquitous 9-kDa protein widely present in the plant kingdom
We have developed enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies specific to barley LTP (bLTP) and proposed its application in quality controlling of beer production
This study introduces the sandwich ELISA using monoclonal antibodies (mAbs) specific to bLTP as a useful tool for quality control and authenticity assessment of beer
Summary
Non-specific lipid transfer protein (LTP) is an ubiquitous 9-kDa protein widely present in the plant kingdom. It is known that LTP is synthesized as a precursor with a typical N-terminal signal peptide and the mature LTP is secreted outside the cell [1,3,4] The former idea about lipid transportation by LTP has been abandoned. The highest expression of LTP has been found in peripheral cell layers surrounding aerial organs, associated with the cell wall and cuticle of epidermal tissues. LTP is composed of eight cysteine residues forming a network of four disulfide bridges These disulfide bonds provide stability in the structure of LTP to heat treatment and proteolytic digestion, which is the characteristic feature of food allergen. Enzyme-linked immunosorbent assay (ELISA) is well known, and is widely utilized as a method for determining the presence of specific proteins in processed foods [26]. We have developed ELISA with monoclonal antibodies (mAbs) specific to bLTP and proposed its application in quality controlling of beer production
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