Table 1.
Checklist of questions to guide experimental design when characterizing the ENM biocorona.
| Experimental design checklist for biocorona characterization |
|---|
| Phase 1: ENM exposure to biofluids and ENM characterization |
| How can the biofluid be handled and stored to best reflect biological conditions? |
| What is the most relevant and appropriate dosage of ENMs in the chosen biofluid or hypothesis? |
| Does the study aim for analysis of the biocorona pre- or post-equilibrium? How was the timeline for equilibrium evaluated? |
| Are replicates and controls included? And, where possible, do plans include parallel processing of samples for proteomics and additional characterisation? |
| Phase 2: Isolation of absorbed biomolecules and preparation for analysis |
| Is the biocorona separation thorough enough to remove all unbound biomolecules? |
| Is the separation technique likely to alter the chemical structure of the biocorona, or the profile of biomolecules? |
| Will the biocorona clean-up steps affect the profile of your biomolecules? |
| What clean-up methods are likely to least alter the biocorona, e.g. through selective or pervasive protein loss? |
| Phase 3: Separation of biomolecules and spectroscopic characterization |
| What type of separation will be best for these samples (in gel, on-particle etc.)? |
| How can the protocol be modified to best maintain consistency across samples? |
| What instruments are best for these samples and quantification? |
| What controls can be included to check for |
| Phase 4: Informatic identification of the biocorona population and analysis. |
| What informatics database will be used for identifying the biomolecules? |
| What kind of post-processing is required for quantification? |
| What statistical analysis of the dataset can be used to assess data quality? |
| How will the full dataset of biomolecules and their characteristics be organized, analysed, and presented? |