Table 2.
Example of Quality-control assays for clinical-grade SB plasmid
Testing | Example assays | Criteria | |
---|---|---|---|
Purity | Identity | Restriction mapping and agarose gel | Expected band size |
Sequencing | Sanger, high through put | Confirm the original coding sequence | |
Concentration | absorbance at 260nm | 2.0 ± 0.2 mg mL−1 | |
A260/A280 absorbance | assay name | 1.8 ± 0.2 | |
Plasmid Form | HPLC | >90% supercoiled | |
Safety | Sterility Test | USP, no growth within 14 days | No growth for 14 days |
Bacterial endotoxin | Kinetic LAL test | <50 EU mg−1 | |
E. coli host protein | ELISA | <0.3% | |
E. coli RNA | HPLC | <10% | |
Others | Appearance | Observation | Clear, colorless, liquid |
Abbreviations: E. coli, Escherichia coli; ELISA, enzyme-linked immunosorbent assay; HPLC, high-performance liquid chromatography; SB, Sleeping Beauty.