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Identification of an Escherichia coli Protein Impurity in Preparations of a Recombinant Pharmaceutical

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Abstract

A host-cell protein impurity found in preparations of recombinant human acidic fibroblast growth factor (aFGF) was identified. Samples of aFGF examined by western blot analysis employing antiserum raised against an Escherichia coli cell lysate contained an immunoreactive protein with a molecular weight of approximately 26,000. The impurity was chromatographically isolated and the N-terminal sequence was determined. Comparing the sequence to a protein database provisionally identified the isolated impurity as the S3 ribosomal protein of E. coli. Monoclonal antibodies recognizing three separate epitopes of S3 confirmed the identity of the impurity in western blots of aFGF samples. The monoclonal antibodies were also used to estimate S3 levels in various preparations of aFGF.

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Authors and Affiliations

  1. Department of Analytical Research, Merck Research Laboratories, P.O. Box 2000, Rahway, New Jersey, 07065

    Donald O. O'Keefe & Mark L. Will

  2. Department of Biochemical Process Research and Development, Merck Research Laboratories, West Point, Pennsylvania, 19486

    Peter DePhillips

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  1. Donald O. O'Keefe
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  2. Peter DePhillips
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  3. Mark L. Will
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O'Keefe, D.O., DePhillips, P. & Will, M.L. Identification of an Escherichia coli Protein Impurity in Preparations of a Recombinant Pharmaceutical. Pharm Res 10, 975–979 (1993). https://doi.org/10.1023/A:1018950319965

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