Novel polyclonal antibody-based rapid gold sandwich immunochromatographic strip for detecting the major royal jelly protein 1 (MRJP1) in honey.

Yifan Zhang,Yong Chen,Jinjie Zhang,Xiaohou Wang,Zongyan Cui,Yiting Cai,Lirong Shen,Amitava Mukherjee

doi:10.1371/journal.pone.0212335

Yifan Zhang, Yong Chen + Show 6 more

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https://doi.org/10.1371/journal.pone.0212335

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Abstract

Honey adulteration is becoming increasingly alarming incidents in food safety. Monitoring and detecting adulteration face greater challenges. Honey contains the major royal jelly proteins (MRJP) secreted by bee workers. To detect honey adulteration fast and accurately, a rapid gold sandwich immunochromatographic strip (GSIS) was developed based on two specific polyclonal antibodies (PoAbs) against the MRJP1, the most abundant protein of all MRJPs. We determined the best of pH value (pH 8.6) and PoAb SP-1 amount (5 μg/mL) in conjunction with colloidal. The cut-off value (sensitivity) of GSIS in detecting MRJP1 is 2.0 μg/mL in solution. Validation analysis with RJ, milk vetch honey, acacia honey and honey adulteration containing rice syrup and corn syrup with different ratios demonstrated that the GSIS could show consistent Test line (T line) when the test samples contain more than 30% pure honey or MRJP1 0.4 mg/g. The validation results by isotope ratio mass spectrometry on the same pure and all adulteration milk vetch honey samples showed the same information of GSIS test. The qualitative assay GSIS provided a valuable new way for honey authenticity and laid the foundation for the future application of GSIS with monoclonal antibodies in honey authentication.

Highlights

Honey is produced by honeybees from the nectar of plants or honeydew secretions of plants combined with their own secretions
The optical density (OD) values of both antibody at 80000-fold dilution were 6.73 and 7.71 times that of the unimmunized rabbit serum (OD at 1000-fold dilution), respectively. This suggests that both antibodies possess very high binding activities to MRJP1, indicating they met the standard of titer for immunological experiments
MRJP1 could be detected in the pre-treated honey samples by the gold sandwich immunochromatographic strip (GSIS)

Summary

Introduction

Honey is produced by honeybees from the nectar of plants or honeydew secretions of plants combined with their own secretions. It is mainly composed of sugar, mainly fructose and glucose, organic acids, amino acids, proteins, minerals, and other phyto-chemicals [1]. The composition of honey depends on its geographical and botanical origins. Honey exhibit a wide range of nutritional and biological effects such as antibacterial property, antioxidant property, antimutagenic property, anti-inflammatory and healing property, etc [2]. As a nutrient-rich natural sweetener and healthcare product, honey is one of the global.

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