title: A non-isotopic probe-hybridization assay for residual DNA in biopharmaceuticals.
abstract: Although most biopharmaceuticals are highly purified, there is a theoretical concern that such recombinant products could be contaminated with oncogenic or bacterial DNA. A crucial part of the control of such biologicals is to ensure they do not contain more residual DNA than a safety limit suggested by the regulatory agency. Currently, the FDA has suggested a 100 pg per dose limit for residual DNA. DNA probes labeled with a radioisotope such as 32P have been commonly used in hybridization tests. Because of the radiation safety concern, we chose to develop a procedure for assessing DNA levels by either a dot or slot blot hybridization technique using a nonisotopic DNA probe and immuno-enzymatic detection. A minimum detectable limit (MDL) of < 10 pg DNA mg(-1) protein can be achieved. Method validation data demonstrated that the precision, reproducibility, and robustness of this approach are appropriate for quality control.
authors.affiliation: Biopharmaceutical Product Development, Lilly Research Laboratories, Lilly Corporate Center, Indianapolis, IN 46285-3848, USA.
authors.forename: A
authors.initials: A
authors.lastname: Riggin
authors.forename: V T
authors.initials: VT
authors.lastname: Luu
authors.forename: J K
authors.initials: JK
authors.lastname: Lobdell
authors.forename: M K
authors.initials: MK
authors.lastname: Wind
mesh_terms: D001702:Biopharmaceutics / Q000592:standards*; D015139:Blotting, Southern; D004247:DNA / Q000592:standards; D015342:DNA Probes / Q000737:chemistry*; D004269:DNA, Bacterial / Q000032:analysis*; D004926:Escherichia coli / Q000235:genetics*; D007004:Hypoglycemic Agents / Q000032:analysis*; D007124:Immunoenzyme Techniques; D007328:Insulin / Q000031:analogs & derivatives* / Q000032:analysis; D061268:Insulin Lispro; D009693:Nucleic Acid Hybridization / Q000379:methods*; D011786:Quality Control; D011994:Recombinant Proteins / Q000032:analysis; D012015:Reference Standards; D015203:Reproducibility of Results
chemical_list: D015342:DNA Probes; D004269:DNA, Bacterial; D007004:Hypoglycemic Agents; D007328:Insulin; D061268:Insulin Lispro; D011994:Recombinant Proteins; D004247:DNA