Abstract
Hens were intramuscularly immunized and rabbits were subcutaneously immunized once every two weeks for 6 weeks using bovine lactoferrin (LF) as antigen. Antibody titers of both yolk (IgY) and rabbit serum (IgG) were as high as 1.68×108 at the 6th and 8th weeks, respectively, after the initial immunization treatment. However, antibody titer against LF in yolk was 9.4×107 at 16 weeks. While antibody titer of rabbit serum declined sharply to 2.1×107 at the 12th week and to 2.6×106 at the 13th week after the initial immunization. The purification efficiency (specific activity of purified antibody against LF/specific activity of the corresponding antiserum or yolk against LF) of rabbit serum IgG purified by laboratory-prepared LF-Sepharose 4B immunoaffinity column (0.05 mg LF/ml wet gel) was about 2400, similar to that of IgY purified by LF-Sepharose 4B immunoaffinity column. Different amounts (0–15.0 mg) of IgY purified by LF-Sepharose 4B immunoaffinity chromatography were applied to the same column to determine the binding capacity (qm) and dissociation constant (Kd) of LF-Sepharose 4B immunoaffinity gel for IgY specific against LF. It was found that qm was 0.81 mg IgY/ml wet gel (1.620 mg IgY/mg LF) and Kd was 6.4×10−6 M as determined by Langmuir-type adsorption isotherms.
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