novel real time polymerase chain reaction approach for rapid detection of the residual escherichia coli genomic dna in biopharmaceutical products establishment of real time polymerase chain reaction to detect residual gdna

نویسندگان

taghi naserpour farivar cellular and molecular research center, qazvin university of medical sciences, qazvin, ir iran; department of clinical biochemistry and genetics, qazvin university of medical sciences, qazvin, ir iran

babak mamnoon cellular and molecular research center, qazvin university of medical sciences, qazvin, ir iran

mohsen karimi arzenani department of molecular medicine, pasteur institute of iran, tehran, ir iran

dariush ilghari department of clinical biochemistry and genetics, qazvin university of medical sciences, qazvin, ir iran; department of clinical biochemistry and genetics, qazvin university of medical sciences, qazvin, ir iran. fax: +98-2833331007

چکیده

background contamination of therapeutic recombinant proteins with residual host cell dna must be controlled under the regulatory standards. objectives the current study established a new rapid, sensitive real time polymerase chain reaction (pcr) approach to measure the reliably of the residual escherichia coli (e. coli) host cell genomic dna in the recombinant streptokinase and alfa interferon preparations. materials and methods in this assay, a specific primer pair was utilized to amplify a 115 base pair sequence inside the e. coli 16s rrna using sybr green chemistry. this method enabled the authors to detect a very small quantity of the residual genomic dna, as low as 0.8 pg, in the protein-based drugs. this method can, therefore, offer a dependable way to quantitatively analyze the major contaminant of biopharmaceutical products, the host cell dna, during the manufacturing process. results sybr green pcr master mix may contain a source of dna contamination during its manufacturing process. conclusions the current study data showed that e. coli host cell dna contamination in streptokinase and alfa interferon manufactured in the pasteur institute of iran is much lower than the safety limits suggested by the fda.

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عنوان ژورنال:
journal of biotechnology and health sciences

جلد ۱، شماره ۳، صفحات ۰-۰

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