title: [A PCR-based semiquantitative assay of DNA impurities in recombinant protein preparations].
abstract: A semiquantitative assay of DNA impurities in preparations of human recombinant insulin is described. The assay is based on the detection of a fragment of the ampicillin-resistant gene within the producer strain DNA by PCR. The analysis of PCR products of the studied preparations and PCR products containing known amounts of E. coli total DNA enabled a quantitative determination of the producer strain DNA content in the preparations under study. The sensitivity of the method is 7 pg of E. coli DNA per 10 microg of human recombinant insulin. The high sensitivity of the method allows us to recommend it for the quantitative determination of DNA content in recombinant preparations that do not inhibit PCR.
authors.forename: A N
authors.initials: AN
authors.lastname: Aleksandrov
authors.forename: Iu S
authors.initials: IuS
authors.lastname: Skoblov
authors.forename: M Iu
authors.initials: MIu
authors.lastname: Skoblov
authors.forename: E D
authors.initials: ED
authors.lastname: Shibanova
authors.forename: D I
authors.initials: DI
authors.lastname: Baĭramashvili
authors.forename: A I
authors.initials: AI
authors.lastname: Miroshnikov
mesh_terms: D004247:DNA / Q000032:analysis* / Q000235:genetics; D006801:Humans; D007328:Insulin / Q000737:chemistry*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry; D012680:Sensitivity and Specificity; D001618:beta-Lactamases / Q000235:genetics*
chemical_list: D007328:Insulin; D011994:Recombinant Proteins; D004247:DNA; D001618:beta-Lactamases