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Cationic polyethyleneimine (PEI)-conjugated gold nanoparticles (AuNPs) that are chemically and physically stable under physiological conditions are an ideal candidate for certain bio-medical applications, in particular DNA transfection. However, the issue remains in reproducibly generating uniform stable species, which can cause the inadequate characterization of the resulting product under relevant conditions and timepoints. The principal objective of the present study was to develop an optimized and reproducible synthetic route for preparing stable PEI-conjugated AuNPs (Au-PEIs). To achieve this objective, a parallel multi-parametric approach involving a total of 96 reaction studies evaluated the importance of 6 key factors: PEI molar mass, PEI structure, molar ratio of PEI/Au, concentration of reaction mixtures, reaction temperature, and reaction time. Application of optimized conditions exhibited narrow size distributions with characteristic surface plasmon resonance absorption and positive surface charge. The optimized Au-PEI product generated by this study exhibits exceptional stability under a physiological isotonic medium (phosphate-buffered saline) over 48 h and shelf-life in ambient condition without any significant change or sedimentation for at least 6 months. Furthermore, the optimized Au-PEI product was highly reproducible. Contributions from individual factors were elucidated using a broad and orthogonal characterization suite examining size and size distribution, optical absorbance, morphological transformation (agglomeration/aggregation), surface functionalities, and stability. Overall, this comprehensive multi-parametric investigation, supported by thorough characterization and rigorous testing, provides a robust foundation for the nanomedicine research community to better synthesize nanomaterials for biomedical use.

journal of nanoparticle research

Parallel multi-parameter study of PEI-functionalized gold nanoparticle synthesis for bio-medical applications: part 1—a critical assessment of methodology, properties, and stability