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Figure 4: Results of representative IgE Immunoblots from samples of standard egg and milk. Picture A was the positive control of rhLF and hLF using specific antibody to rhLF and hLF. The order of the laoded samples in picture A were Lane 1: recombinant human lactoferrin (rhLF, 40 ng), Lane 2: human lactoferrin (hLF, 40 ng), Lane 3: ddH2O (15 μl). The order of the laoded samples in the picture B was as Lane 1: standard milk powder extracts (10 μg), Lane 2: standard egg powder extracts (10 μg), Lane 3: recombinant human lactoferrin (rhLF, 40 ng), Lane 4: human lactoferrin (hLF, 40 ng), Lane 5: bovine lactoferrin (bLF, 40 ng). The serum used in picture B to incubate the membrane was mixture of 5 control sera. In picture C and D, Lane 1: standard egg powder extracts in C (10 μg) or standard milk powder extracts in D (10 μg), Lane 2: recombinant human lactoferrin (rhLF, 40 ng), Lane 3: human lactoferrin (hLF, 40 ng), Lane 4: bovine lactoferrin (bLF, 40 ng). The C was obtained by incubating the memnrane with a pool contained 12 sera from individual donors allergic to egg. The level of egg specific IgE in all of the 12 sera samples were >3.5 KUA/L. Finally, 21 sera donated from individual donors allergic to milk were mixed to incubate the membrane D, and the level of milk specific IgE in the serum was also >3.5 KUA/L. In egg and milk there were sevral proteins binded to specific allergic serum, while the rhLF, hLF and bLF. This indecate that there was not cross reaction between rhLF and egg, milk.
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