PMC4293047_01
SDS-PAGE and Western blot results support casein-specific allergy. Casein and whey (alpha-lactalbumin and beta-lactoglobulin) are the main proteins in CM (column CM). pHWF has no casein proteins (column pHWF).The case patient's serum showed binding to only casein proteins in CM (column Pt-pHWF), and no binding to any proteins in pHWF(column Pt-pHWF). Pooled sera from subjects with typical CM allergy showed binding to casein and whey proteins in CM (column pool-CM) and to whey proteins and residual casein in pHWF(column pool-pHWF). The negative control showed no binding.Pt= patient, CM=cow's milk, pHWF=partially hydrolyzed whey formula, NC=negative control, alpha-La=alpha-lactalbumin, beta-Lg=beta-lactoglobulin
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F1
ImmobilonP (Figure 1, left panel). CM showed proteins at 14 kDa, 18 kDa, and >28kDa, corresponding to alpha-lactalbumin, beta-lactoglobulin, and casein proteins, respectively. Irrespective of the quantity used (25-150 mug), pHWF showed only very low molecular weight hydrolyzed product (3-14 kDa). There was evidence of the patient's serum sIgE binding to higher molecular weight proteins in CM (>= 28kDa, corresponding to casein and its aggregates) (Figure 1, column Pt-CM). In contrast, the patient's serum did not show any binding to proteins in pHWF (Figure 1, column Pt-pHWF). The pooled serum showed binding to casein and whey proteins in CM (Figure 1, column Pool-CM). The pooled serum bound to lower molecular weight bands corresponding to whey proteins in pHWF (Figure 1, column Pool-pHWF).