Isolation and characterization of immunoglobulin in yolk (IgY) specific against hen egg white lysozyme by immunoaffinity chromatography.
Six hens were intramuscularly (im) immunized once a week for 3 weeks using chicken egg white lysozyme (LS) as antigen. Antibody (immunoglobulin in yolk, IgY) ELISA values of 10(3)-fold diluted yolk were almost as high as 1.879 in the sixth week and maintained a value of 0.756 in the eighth week after the initial immunization treatment. The purification efficiency (specific activity of purified IgY against LS/specific activity of antibody in yolk against LS) of IgY specific against LS isolated by laboratory-prepared LS-bound (IgY-) Sepharose 4 Fast Flow immunoaffinity column was approximately 3380. By applying various amounts (0-22 mg) of the thusly obtained IgY specific against LS to the immunoaffinity column, the binding capacity (q(m)) and dissociation constant (K(d), M(-1)) of such immunoaffinity gel for IgY against LS were found to be 0.68 mg of IgY/mL of wet gel (0.54 mg of IgY/mg of LS) and 7.13 x 10(-6) M, respectively, as determined by Langmuir-type adsorption isotherms.
Department of Food Health, Chia-Nan University of Pharmacy and Science, Tainan 717, Taiwan.
Chao-Cheng
CC
Chen
Yann-Ying
YY
Tu
Tzy-Li
TL
Chen
Hung-Min
HM
Chang
D000818:Animals; D000918:Antibody Specificity; D002645:Chickens / Q000276:immunology*; D004529:Egg White / Q000032:analysis*; D004530:Egg Yolk / Q000276:immunology*; D007114:Immunization; D007136:Immunoglobulins / Q000276:immunology / Q000302:isolation & purification*; D009113:Muramidase / Q000276:immunology*
C016512:IgY; D007136:Immunoglobulins; D009113:Muramidase