Combining 2-DE immunoblots and mass spectrometry to identify putative soybean (Glycine max) allergens

Mei Lu, Yuan Jin, Ronald Cerny, Barbara Ballmer-Weber, Richard E Goodman

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Soybean is recognized as a commonly allergenic food, but the identity of important allergens is not well studied. Recently, some global regulatory agencies started requiring quantitative analysis of individual allergens, including unproven allergens, as part of the risk assessment for genetically engineered (GE) soybeans. We sought to identify soybean proteins that bind IgE from any of 10 individual soybean-sensitized subjects. Soybean IgE binding proteins were identified by 2-DE immunoblots using sera from four soy-allergic and plasma from six soy-sensitized human subjects. Corresponding spots were excised from stained gels, digested, and analyzed using a quadrupole TOF Synapt G2-S tandem mass spectrometer. Results showed the major IgE binding proteins were subunits of either β-conglycinin (Gly m 5) or glycinin (Gly m 6). Soybean Kunitz trypsin inhibitor (SKTI) was a significant IgE binding protein for four subjects. Soybean agglutinin, seed biotinylated protein (SBP) of 65 kDa, late embryogenesis protein (LEP), and sucrose-binding protein were identified as IgE binding only for soy-sensitized subjects. We conclude that the major soybean allergens are isoforms of Gly m 5, Gly m 6, and possibly SKTI and that requirements for quantitative measurement of proteins that are not clear allergens is not relevant to safety.

Original languageEnglish (US)
Pages (from-to)207-215
Number of pages9
JournalFood and Chemical Toxicology
Volume116
DOIs
StatePublished - Jun 1 2018

Fingerprint

allergens
Soybeans
Allergens
Glycine
Mass spectrometry
Glycine max
Mass Spectrometry
Galectin 3
mass spectrometry
soybeans
Kunitz Soybean Trypsin Inhibitor
binding proteins
Immunoglobulin E
trypsin inhibitors
soy protein
Soybean Proteins
Proteins
Protein Subunits
Mass spectrometers
Protein Binding

Keywords

  • Glycinin
  • IgE
  • LC-MS/MS
  • Soybean allergens
  • Two-dimensional immunoblots
  • β-conglycinin

ASJC Scopus subject areas

  • Food Science
  • Toxicology

Cite this

Combining 2-DE immunoblots and mass spectrometry to identify putative soybean (Glycine max) allergens. / Lu, Mei; Jin, Yuan; Cerny, Ronald; Ballmer-Weber, Barbara; Goodman, Richard E.

In: Food and Chemical Toxicology, Vol. 116, 01.06.2018, p. 207-215.

Research output: Contribution to journalArticle

@article{4ded52c808e745b79c464793a7e56a8a,
title = "Combining 2-DE immunoblots and mass spectrometry to identify putative soybean (Glycine max) allergens",
abstract = "Soybean is recognized as a commonly allergenic food, but the identity of important allergens is not well studied. Recently, some global regulatory agencies started requiring quantitative analysis of individual allergens, including unproven allergens, as part of the risk assessment for genetically engineered (GE) soybeans. We sought to identify soybean proteins that bind IgE from any of 10 individual soybean-sensitized subjects. Soybean IgE binding proteins were identified by 2-DE immunoblots using sera from four soy-allergic and plasma from six soy-sensitized human subjects. Corresponding spots were excised from stained gels, digested, and analyzed using a quadrupole TOF Synapt G2-S tandem mass spectrometer. Results showed the major IgE binding proteins were subunits of either β-conglycinin (Gly m 5) or glycinin (Gly m 6). Soybean Kunitz trypsin inhibitor (SKTI) was a significant IgE binding protein for four subjects. Soybean agglutinin, seed biotinylated protein (SBP) of 65 kDa, late embryogenesis protein (LEP), and sucrose-binding protein were identified as IgE binding only for soy-sensitized subjects. We conclude that the major soybean allergens are isoforms of Gly m 5, Gly m 6, and possibly SKTI and that requirements for quantitative measurement of proteins that are not clear allergens is not relevant to safety.",
keywords = "Glycinin, IgE, LC-MS/MS, Soybean allergens, Two-dimensional immunoblots, β-conglycinin",
author = "Mei Lu and Yuan Jin and Ronald Cerny and Barbara Ballmer-Weber and Goodman, {Richard E}",
year = "2018",
month = "6",
day = "1",
doi = "10.1016/j.fct.2018.04.032",
language = "English (US)",
volume = "116",
pages = "207--215",
journal = "Food and Chemical Toxicology",
issn = "0278-6915",
publisher = "Elsevier Limited",

}

TY - JOUR

T1 - Combining 2-DE immunoblots and mass spectrometry to identify putative soybean (Glycine max) allergens

AU - Lu, Mei

AU - Jin, Yuan

AU - Cerny, Ronald

AU - Ballmer-Weber, Barbara

AU - Goodman, Richard E

PY - 2018/6/1

Y1 - 2018/6/1

N2 - Soybean is recognized as a commonly allergenic food, but the identity of important allergens is not well studied. Recently, some global regulatory agencies started requiring quantitative analysis of individual allergens, including unproven allergens, as part of the risk assessment for genetically engineered (GE) soybeans. We sought to identify soybean proteins that bind IgE from any of 10 individual soybean-sensitized subjects. Soybean IgE binding proteins were identified by 2-DE immunoblots using sera from four soy-allergic and plasma from six soy-sensitized human subjects. Corresponding spots were excised from stained gels, digested, and analyzed using a quadrupole TOF Synapt G2-S tandem mass spectrometer. Results showed the major IgE binding proteins were subunits of either β-conglycinin (Gly m 5) or glycinin (Gly m 6). Soybean Kunitz trypsin inhibitor (SKTI) was a significant IgE binding protein for four subjects. Soybean agglutinin, seed biotinylated protein (SBP) of 65 kDa, late embryogenesis protein (LEP), and sucrose-binding protein were identified as IgE binding only for soy-sensitized subjects. We conclude that the major soybean allergens are isoforms of Gly m 5, Gly m 6, and possibly SKTI and that requirements for quantitative measurement of proteins that are not clear allergens is not relevant to safety.

AB - Soybean is recognized as a commonly allergenic food, but the identity of important allergens is not well studied. Recently, some global regulatory agencies started requiring quantitative analysis of individual allergens, including unproven allergens, as part of the risk assessment for genetically engineered (GE) soybeans. We sought to identify soybean proteins that bind IgE from any of 10 individual soybean-sensitized subjects. Soybean IgE binding proteins were identified by 2-DE immunoblots using sera from four soy-allergic and plasma from six soy-sensitized human subjects. Corresponding spots were excised from stained gels, digested, and analyzed using a quadrupole TOF Synapt G2-S tandem mass spectrometer. Results showed the major IgE binding proteins were subunits of either β-conglycinin (Gly m 5) or glycinin (Gly m 6). Soybean Kunitz trypsin inhibitor (SKTI) was a significant IgE binding protein for four subjects. Soybean agglutinin, seed biotinylated protein (SBP) of 65 kDa, late embryogenesis protein (LEP), and sucrose-binding protein were identified as IgE binding only for soy-sensitized subjects. We conclude that the major soybean allergens are isoforms of Gly m 5, Gly m 6, and possibly SKTI and that requirements for quantitative measurement of proteins that are not clear allergens is not relevant to safety.

KW - Glycinin

KW - IgE

KW - LC-MS/MS

KW - Soybean allergens

KW - Two-dimensional immunoblots

KW - β-conglycinin

UR - http://www.scopus.com/inward/record.url?scp=85045769581&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85045769581&partnerID=8YFLogxK

U2 - 10.1016/j.fct.2018.04.032

DO - 10.1016/j.fct.2018.04.032

M3 - Article

VL - 116

SP - 207

EP - 215

JO - Food and Chemical Toxicology

JF - Food and Chemical Toxicology

SN - 0278-6915

ER -