Isolation and characterization of immunoglobulin in yolk (IgY) specific against hen egg white lysozyme by immunoaffinity chromatography

J Agric Food Chem. 2002 Sep 11;50(19):5424-8. doi: 10.1021/jf011567h.

Abstract

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.

MeSH terms

  • Animals
  • Antibody Specificity*
  • Chickens / immunology*
  • Egg White / analysis*
  • Egg Yolk / immunology*
  • Immunization
  • Immunoglobulins / immunology
  • Immunoglobulins / isolation & purification*
  • Muramidase / immunology*

Substances

  • IgY
  • Immunoglobulins
  • Muramidase