J. Anim Sci.
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J. Anim. Sci. 2002. 80:1863-1870
© 2002 American Society of Animal Science

Purification of porcine ß-casein, N-terminal sequence, quantification in mastitic milk1,2

A. C. W. Kauf and R. S. Kensinger3

Department of Dairy and Animal Science, Pennsylvania State University, University Park 16802

3 Correspondence:
phone: 814-863-0558; fax: 814-865-7442; E-mail:
RSK7{at}psu.edu.

The objectives of the study were to purify porcine ß-casein from sow’s milk, to determine N-terminal amino acid sequence, to develop specific antisera against porcine ß-casein, and to use that antisera to evaluate milk samples from a mastitis study. Milk was collected by hand milking a Yorkshire by Duroc crossbred sow following oxytocin administration on d 27 of lactation. A casein-enriched fraction was then prepared by iso-electric precipitation. Porcine ß-casein was then purified by liquid chromatography on a Mono Q anion-exchange column, and checked for purity with SDS-PAGE. An apparent molecular weight of 29,000 Da was estimated from SDS-PAGE. N-Terminal amino acid sequence was determined by Edman degradation to be RAKEELNASGETVE. Rabbits (n = 2) were immunized with ß-casein mixed with Freund’s complete (primary) or incomplete (boosters) adjuvant at 4-wk intervals. Antiserum collected from one rabbit 112 d after primary immunization detected 30 to 100 ng ß-casein by Western blot procedure when used at a dilution of 1:2 x 106. The antiserum was specific for porcine ß-casein, but showed some cross-reactivity with equine casein. It was determined by Western blot procedure that mammary inflammation induced by lipopolysaccharide infusion resulted in a 41% decrease in the ß-casein concentration of sow milk.

Key Words: Amino Acid Sequences • Antiserum • ß-Casein • Liquid Chromatography • Mastitis • Pigs






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