Purification of recombinant lecithin:cholesterol acyltransferase

Maya P. Nair, Bhalchandra J. Kudchodkar, P. Haydn Pritchard, Andras G. Lacko

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Production and purification of recombinant human lecithin:cholesterol acyltransferase (LCAT), secreted by baby hamster kidney (BHK) cells, has been improved by limiting the harvesting times for the conditioned medium and introducing an additional purification step. The recombinant BHK cells were grown until nearly confluent on multilayered flasks in a fetal-calf-serum-enriched medium. Subsequently, the cells were washed and supplied with serum free medium for 24-h periods. The conditioned medium, containing recombinant LCAT, was harvested at 24 and 48 h and subjected to chromatography on phenyl-Sepharose and ACA-44 agarose to isolate the recombinant enzyme. The second chromatography step revealed the presence of a low-molecular-weight contaminant that exhibited a carbohydrate/protein composition similar to proteoglycans. The major purified component contained LCAT activity and was homogeneous by acrylamide gel electrophoresis.

Original languageEnglish
Pages (from-to)38-41
Number of pages4
JournalProtein Expression and Purification
Volume10
Issue number1
DOIs
StatePublished - 1 Jan 1997

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Phosphatidylcholine-Sterol O-Acyltransferase
Conditioned Culture Medium
Cricetinae
Chromatography
Kidney
Acrylamide
Serum-Free Culture Media
Proteoglycans
Sepharose
Electrophoresis
Molecular Weight
Gels
Carbohydrates
Enzymes
Serum
Proteins

Cite this

Nair, Maya P. ; Kudchodkar, Bhalchandra J. ; Pritchard, P. Haydn ; Lacko, Andras G. / Purification of recombinant lecithin:cholesterol acyltransferase. In: Protein Expression and Purification. 1997 ; Vol. 10, No. 1. pp. 38-41.
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Purification of recombinant lecithin:cholesterol acyltransferase. / Nair, Maya P.; Kudchodkar, Bhalchandra J.; Pritchard, P. Haydn; Lacko, Andras G.

In: Protein Expression and Purification, Vol. 10, No. 1, 01.01.1997, p. 38-41.

Research output: Contribution to journalArticle

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