A non-isotopic probe-hybridization assay for residual DNA in biopharmaceuticals

• Published in: Journal of pharmaceutical and biomedical analysis Vol. 16; no. 4; pp. 561 - 572
• Main Authors: Riggin, A, Luu, V.T, Lobdell, J.K, Wind, M.K
• Format: Journal Article Conference Proceeding
• Language: English
• Published: Amsterdam Elsevier B.V 01.12.1997; Elsevier Science
• Subjects: Analysis; Biological and medical sciences; Biopharmaceuticals; Biopharmaceutics - standards; Blotting, Southern; DNA - standards; DNA Probes - chemistry; DNA, Bacterial - analysis; Dot-blot/slot-blot technique; Escherichia coli - genetics; General pharmacology; Hypoglycemic Agents - analysis; Immunoenzyme Techniques; Insulin - analogs & derivatives; Insulin - analysis; Insulin Lispro; Medical sciences; Non-isotopic assay; Nucleic Acid Hybridization - methods; Pharmacology. Drug treatments; Probe hybridization assay; Quality Control; Recombinant Proteins - analysis; Reference Standards; Reproducibility of Results; Residual DNA detection; Sulfonated DNA; Non-isotopic assay; Biopharmaceuticals; Dot-blot/slot-blot technique; Probe hybridization assay; Residual DNA detection; Sulfonated DNA; Drug; Validation; Purity control; Hybridization; Enzyme immunoassay; In vitro; DNA; Detection limit; Quality control; Bacteria; Molecular probe; Recombinant protein; Radiopharmaceuticals; Biological contamination; Quantitative analysis
• ISSN: 0731-7085; 1873-264X
• DOI: 10.1016/S0731-7085(97)00152-0

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.