Isolation of recombinant proteins from milk

Tracy D. Wilkins, William Velander

Research output: Contribution to journalArticle

29 Citations (Scopus)

Abstract

Milk is a complex bio‐colloid which presents some unique problems for the protein isolation chemist, but the majority of the processing criteria for purifying recombinant proteins are the same as with any complex biological mixture. The casein micelles and fat globules behave as separate phases; they prevent filtration of the milk and interfere with the usual separation methods. The usual first step is to centrifuge the milk to remove the fat and precipitate the casein micelles with low pH or precipitating agents. Some recombinant proteins may associate to some degree with the micelles which may necessitate solubilizing them with chelating agents. If the majority of the product protein associates with either the fat or micelles, this can be used to advantage. Once the casein micelles have been removed or disrupted, the clarified milk can be processed by the usual separation methods. There also are proteases in milk which can degrade recombinant proteins. The greatest advantage of producing recombinant proteins in milk is the high concentration which can be obtained. The high levels of product protein can alleviate many problems associated with the application of classical purification strategies to transgenic milk proteins.

Original languageEnglish (US)
Pages (from-to)333-338
Number of pages6
JournalJournal of Cellular Biochemistry
Volume49
Issue number4
DOIs
StatePublished - Aug 1992

Fingerprint

Recombinant Proteins
Micelles
Milk
Caseins
Fats
Proteins
Milk Proteins
Centrifuges
Chelating Agents
Complex Mixtures
Purification
Precipitates
Peptide Hydrolases
Processing

Keywords

  • Casein micelle
  • Immunopurification
  • Lactation
  • Monoclonal antibody
  • Post‐translational modification
  • proteases
  • therapeutics
  • viral inactivation
  • whey

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Isolation of recombinant proteins from milk. / Wilkins, Tracy D.; Velander, William.

In: Journal of Cellular Biochemistry, Vol. 49, No. 4, 08.1992, p. 333-338.

Research output: Contribution to journalArticle

Wilkins, Tracy D. ; Velander, William. / Isolation of recombinant proteins from milk. In: Journal of Cellular Biochemistry. 1992 ; Vol. 49, No. 4. pp. 333-338.
@article{39f33f41f33d4893827bec4e41cee4be,
title = "Isolation of recombinant proteins from milk",
abstract = "Milk is a complex bio‐colloid which presents some unique problems for the protein isolation chemist, but the majority of the processing criteria for purifying recombinant proteins are the same as with any complex biological mixture. The casein micelles and fat globules behave as separate phases; they prevent filtration of the milk and interfere with the usual separation methods. The usual first step is to centrifuge the milk to remove the fat and precipitate the casein micelles with low pH or precipitating agents. Some recombinant proteins may associate to some degree with the micelles which may necessitate solubilizing them with chelating agents. If the majority of the product protein associates with either the fat or micelles, this can be used to advantage. Once the casein micelles have been removed or disrupted, the clarified milk can be processed by the usual separation methods. There also are proteases in milk which can degrade recombinant proteins. The greatest advantage of producing recombinant proteins in milk is the high concentration which can be obtained. The high levels of product protein can alleviate many problems associated with the application of classical purification strategies to transgenic milk proteins.",
keywords = "Casein micelle, Immunopurification, Lactation, Monoclonal antibody, Post‐translational modification, proteases, therapeutics, viral inactivation, whey",
author = "Wilkins, {Tracy D.} and William Velander",
year = "1992",
month = "8",
doi = "10.1002/jcb.240490403",
language = "English (US)",
volume = "49",
pages = "333--338",
journal = "Journal of Cellular Biochemistry",
issn = "0730-2312",
publisher = "Wiley-Liss Inc.",
number = "4",

}

TY - JOUR

T1 - Isolation of recombinant proteins from milk

AU - Wilkins, Tracy D.

AU - Velander, William

PY - 1992/8

Y1 - 1992/8

N2 - Milk is a complex bio‐colloid which presents some unique problems for the protein isolation chemist, but the majority of the processing criteria for purifying recombinant proteins are the same as with any complex biological mixture. The casein micelles and fat globules behave as separate phases; they prevent filtration of the milk and interfere with the usual separation methods. The usual first step is to centrifuge the milk to remove the fat and precipitate the casein micelles with low pH or precipitating agents. Some recombinant proteins may associate to some degree with the micelles which may necessitate solubilizing them with chelating agents. If the majority of the product protein associates with either the fat or micelles, this can be used to advantage. Once the casein micelles have been removed or disrupted, the clarified milk can be processed by the usual separation methods. There also are proteases in milk which can degrade recombinant proteins. The greatest advantage of producing recombinant proteins in milk is the high concentration which can be obtained. The high levels of product protein can alleviate many problems associated with the application of classical purification strategies to transgenic milk proteins.

AB - Milk is a complex bio‐colloid which presents some unique problems for the protein isolation chemist, but the majority of the processing criteria for purifying recombinant proteins are the same as with any complex biological mixture. The casein micelles and fat globules behave as separate phases; they prevent filtration of the milk and interfere with the usual separation methods. The usual first step is to centrifuge the milk to remove the fat and precipitate the casein micelles with low pH or precipitating agents. Some recombinant proteins may associate to some degree with the micelles which may necessitate solubilizing them with chelating agents. If the majority of the product protein associates with either the fat or micelles, this can be used to advantage. Once the casein micelles have been removed or disrupted, the clarified milk can be processed by the usual separation methods. There also are proteases in milk which can degrade recombinant proteins. The greatest advantage of producing recombinant proteins in milk is the high concentration which can be obtained. The high levels of product protein can alleviate many problems associated with the application of classical purification strategies to transgenic milk proteins.

KW - Casein micelle

KW - Immunopurification

KW - Lactation

KW - Monoclonal antibody

KW - Post‐translational modification

KW - proteases

KW - therapeutics

KW - viral inactivation

KW - whey

UR - http://www.scopus.com/inward/record.url?scp=0026760444&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026760444&partnerID=8YFLogxK

U2 - 10.1002/jcb.240490403

DO - 10.1002/jcb.240490403

M3 - Article

C2 - 1429861

AN - SCOPUS:0026760444

VL - 49

SP - 333

EP - 338

JO - Journal of Cellular Biochemistry

JF - Journal of Cellular Biochemistry

SN - 0730-2312

IS - 4

ER -