Pediatr Allergy Respir Dis.  1997 Sep;7(2):207-217.

Comparison of Antigenicities of Whole Milk and Partial Hydrolysate of Cow's Milk Proteins by Western Blot Analysis

Affiliations
  • 1Department of Pediatrics, Ajou University School of Medicine, Suwon, Korea.

Abstract

For the prevention or the management of milk allergy in infancy, partial or extensive hydrolysates of cow's milk have been used in western countries for a couple of decades. Recently a Korean product of partial hydrolysate(HA-21) became available for the prevention of sensitization to cow's milk proteins. In this study, to compare the antigenicities of whole milk(WM) and HA-21, we performed the IgG and IgE western-blot analysis. Sera were obtained from 17 milk sensitive infants and 2 controls, and crude extract of WM and HA-21 and purified beta-lactoglobulin(BLG), bovine serum IgG(B-IgG), alpha-lactalbumin(ALA) were used for blot- inhibition study. After the non-reduced SDS-PAGE, western-blot studies were done using biotinylated anti-human IgG and IgE antibodies, and the reaction were detected by avidine-phosphatase system. By the SDS-PAGE analysis, WM were separated into bovine-IgG(B-IgG), bovine serum albumin(BSA), 56 KD protein, caseins(CAS), BLG and ALA, but there was no visible bands above 14 KD in the case of HA-21. In preliminary blot analysis with 4 milk sensitive sera, we found the broad and strong IgE-binding protein band in the range of 150-200 KD in all cases. This fraction, named as 'P-band' in this study, was completely inhibited by BLG and partially inhibited by B-IgG using blot-inhibition study. Using the western blot analysis of WM, the P-band revealed the most prevalent IgG and IgE binding protein in 16 of 17 tested sera. More than 15 bands were identified by IgG-blot of WM, but the IgE biding proteins were P-band(16 sera), BSA(12 sera), 56 KD protein(8 sera), and CAS(5 sera). In the case of HA-21, BSA(8 sera) was the only protein bound with IgE antibody and the reaction was very weak compared to that of WM. In conclusion, the major IgE binding protein appears to be the p-band which might contain the epitopes of B-IgG and BLG in this study. The antigenicity of HA-21 is remarkably reduced compared to WM, but still minute residual antigenicity and allergenicity are remained in the extract of HA-21.

Keyword

Antigenicity; Allergenicity; Whole milk; Partial hydrolysate; Western blot

MeSH Terms

Antibodies
Blotting, Western*
Electrophoresis, Polyacrylamide Gel
Epitopes
Galectin 3
Humans
Immunoglobulin E
Immunoglobulin G
Infant
Milk Hypersensitivity
Milk Proteins*
Milk*
Antibodies
Epitopes
Galectin 3
Immunoglobulin E
Immunoglobulin G
Milk Proteins
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