Non-Invasive Delivery of Negatively Charged Nanobodies by Anodal Iontophoresis: When Electroosmosis Dominates Electromigration

• Published in: Pharmaceutics Vol. 16; no. 4; p. 539
• Main Authors: Sahraoui, Phedra Firdaws, Vadas, Oscar, Kalia, Yogeshvar N
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.04.2024; MDPI
• Subjects: Amino acids; anti-EGFR 7D12 nanobody; Antibodies; Antigens; Electric fields; electroosmosis; Iontophoresis; Molecular weight; Nafarelin; nanobodies; Patient compliance; Peptides; Physiology; protein expression and purification; Proteins; Recombinant proteins; skin topical and transdermal delivery; Germany; United Kingdom > UK; United States > US; Switzerland; nanobodies; electroosmosis; skin topical and transdermal delivery; iontophoresis; protein expression and purification; anti-EGFR 7D12 nanobody
• ISSN: 1999-4923; 1999-4923
• DOI: 10.3390/pharmaceutics16040539

Iontophoresis enables the non-invasive transdermal delivery of moderately-sized proteins and the needle-free cutaneous delivery of antibodies. However, simple descriptors of protein characteristics cannot accurately predict the feasibility of iontophoretic transport. This study investigated the cathodal and anodal iontophoretic transport of the negatively charged M7D12H nanobody and a series of negatively charged variants with single amino acid substitutions. Surprisingly, M7D12H and its variants were only delivered transdermally by anodal iontophoresis. In contrast, transdermal permeation after cathodal iontophoresis and passive diffusion was <LOQ. The anodal iontophoretic delivery of these negatively charged proteins was achieved because electroosmosis was the dominant electrotransport mechanism. Cutaneous deposition after the anodal iontophoresis of M7D12H (wild type), and the R54E and K65E variants, was statistically superior to that after cathodal iontophoresis (6.07 ± 2.11, 9.22 ± 0.80, and 14.45 ± 3.45 μg/cm , versus 1.12 ± 0.30, 0.72 ± 0.27, and 0.46 ± 0.07 µg/cm , respectively). This was not the case for S102E, where cutaneous deposition after anodal and cathodal iontophoresis was 11.89 ± 0.87 and 8.33 ± 2.62 µg/cm , respectively; thus, a single amino acid substitution appeared to be sufficient to impact the iontophoretic transport of a 17.5 kDa protein. Visualization studies using immunofluorescent labeling showed that skin transport of M7D12H was achieved via the intercellular and follicular routes.