Analysis of major milk whey proteins by immunoaffinity capillary electrophoresis coupled with MALDI-MS.
نویسندگان
چکیده
Two major milk whey proteins, β-lactoglobulin and α-lactalbumin, are among the main cow milk allergens and can cause allergy even at a very low concentrations. Therefore, these proteins are interesting targets in food analysis, not only for food quality control but also for highlighting the presence of allergens. Herein, a sensitive analysis for β-lactoglobulin and α-lactalbumin was developed using immunoaffinity capillary electrophoresis hyphenated with MALDI-MS. Magnetic beads functionalized with appropriate antibodies were used for β-lactoglobulin and α-lactalbumin immunocapture inside the capillary. After elution from the beads, analyte focusing and separation were performed by transient isotachophoresis followed by MALDI-MS analysis performed through an automated iontophoretic fraction collection interface. A LOD in the low nanomolar range was attained for both whey proteins. The method developed was further applied to the analysis of different milk samples including fortified soy milk.
منابع مشابه
Component-resolved diagnostic of cow's milk allergy by immunoaffinity capillary electrophoresis-matrix assisted laser desorption/ionization mass spectrometry.
Component-resolved diagnostic (CRD) of cow's milk allergy has been performed using immunoaffinity capillary electrophoresis (IACE) coupled with matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS). First, total IgE quantification in the blood serum of a milk allergic patient by the IACE-UV technique was developed using magnetic beads (MBs) coated with antihuman IgE antibodie...
متن کاملProteomics data in support of the quantification of the changes of bovine milk proteins during mammary gland involution
Here we provide data from three proteomics techniques; two-dimensional electrophoresis (2-DE) followed by identification of selected spots using PSD MALDI-TOF MS/MS, one-dimensional gel electrophoresis followed by LC-MS/MS analysis of gel slices (GeLC) and dimethyl isotopic labelling of tryptic peptides followed by Orbitrap MS/MS (DML), to quantify the changes in the repertoire of bovine milk p...
متن کاملProteomic Analysis of Whey from Bovine Colostrum and Mature Milk
The aim of this study was to standardize a methodology to obtain two-dimensional (2D) maps of whey proteins from bovine colostrum and mature milk, using two-dimensional electrophoresis, in order to identify the minor proteins by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI TOF/TOF MS). A total of 38 proteins were identified, 20 spots in the colostru...
متن کاملThe multiplicity of N-glycan structures of bovine milk 18 kda lactophorin (milk GlyCAM-1).
Lactophorin is a heat-stable phosphoglycoprotein, also known as milk glycosylation-dependent cell adhesion molecule 1 (GlyCAM-1). Bovine 18 kDa lactophorin was purified by heparin affinity chromatography from cow's milk whey. Its N-glycans were obtained by proteomic techniques, including two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), followed by in-gel digestion with peptide-N(4)...
متن کاملAnalysis of bovine whey proteins in soybean dairy-like products by capillary electrophoresis.
The simultaneous separation of bovine whey proteins [alpha-lactalbumin and beta-lactoglobulin (A+B)] and soybean proteins was performed, for the first time, by capillary electrophoresis. Different experimental conditions were tested. The most suitable consisted of 0.050 M phosphate buffer (pH 8) with 1 M urea and 1.2 mg/ml methylhydroxyethylcellulose, UV detection at 280 nm, 15 kV applied volta...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- Electrophoresis
دوره 33 15 شماره
صفحات -
تاریخ انتشار 2012