Milk Alkaline Phosphatase Purification and Production of Polyclonal Antibodies

Authors

  • A. V. Vega-Warner,

    1. Authors Vega-Warner, Smith and Ustunol are affiliated with the Dept. of Food Science & Human Nutrition, Michigan State Univ., E. Lansing, MI 48824.
    Search for more papers by this author
  • C.-H. Wang,

    1. Authors Vega-Warner, Smith and Ustunol are affiliated with the Dept. of Food Science & Human Nutrition, Michigan State Univ., E. Lansing, MI 48824.
    Search for more papers by this author
  • D. M. Smith,

    1. Authors Vega-Warner, Smith and Ustunol are affiliated with the Dept. of Food Science & Human Nutrition, Michigan State Univ., E. Lansing, MI 48824.
    Search for more papers by this author
  • Z. Ustunol

    Corresponding author
    1. Authors Vega-Warner, Smith and Ustunol are affiliated with the Dept. of Food Science & Human Nutrition, Michigan State Univ., E. Lansing, MI 48824.
    Search for more papers by this author

  • Author Wang's current address: Dept. of Food & Nutrition, Providence Univ., Shalu, Taiwan.

  • The financial support of the Michigan Agricultural Experiment Station and State of Michigan Crop and Food Bioprocessing Center is gratefully acknowledged.

Address inquiries to Dr. Z. Ustunol.

ABSTRACT

Purification was by electroelution from native polyacrylamide gels or by sequential use of three columns. Electroelution was faster and resulted in a higher yield (23.4 vs 1.6%) than column purification. The enzyme had a molecular mass of 187 kDa, and the isoelectric point ranged from 5.4 to 6.0. ALP purified by electroelution was used as the antigen to immunize rabbits for polyclonal antibody (PAb) production. Western blot analysis showed that PAbs cross-reacted with bovine milk and placenta ALP, but did not cross-react with ALP from calf or bovine intestinal mucosa, Escherichia coli or with other milk proteins.

Ancillary