Comparative Evaluation of Engineered Polypeptide Scaffolds in HER2-Targeting Magnetic Nanocarrier Delivery

Victoria O Shipunova; Olga A Kolesnikova; Polina A Kotelnikova; Vladislav D Soloviev; Anton A Popov; Galina M Proshkina; Maxim P Nikitin; Sergey M Deyev

doi:10.1021/acsomega.1c01811

Comparative Evaluation of Engineered Polypeptide Scaffolds in HER2-Targeting Magnetic Nanocarrier Delivery

ACS Omega. 2021 Jun 10;6(24):16000-16008. doi: 10.1021/acsomega.1c01811. eCollection 2021 Jun 22.

Authors

Victoria O Shipunova^{1

2

3

4}, Olga A Kolesnikova¹, Polina A Kotelnikova¹, Vladislav D Soloviev^{1

4}, Anton A Popov³, Galina M Proshkina¹, Maxim P Nikitin^{1

2

4}, Sergey M Deyev^{1

3}

Affiliations

¹ Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 16/10 Miklukho-Maklaya Street, Moscow 117997, Russia.
² Moscow Institute of Physics and Technology, 9 Institutskiy per., Dolgoprudny 141701, Russia.
³ MEPhI (Moscow Engineering Physics Institute), Institute of Engineering Physics for Biomedicine (PhysBio), 31 Kashirskoe Shosse, Moscow 115409, Russia.
⁴ Sirius University of Science and Technology, 1 Olympic Avenue, Sochi 354340, Russia.

Abstract

Targeted drug delivery is one of the most intriguing and challenging issues in modern biomedicine. For active targeting, full-size IgG molecules (150 kDa) are usually used. Recent studies have revealed that small artificial polypeptide scaffolds such as DARPins (14 kDa) and affibodies (8 kDa) are much more promising tools for drug delivery due to their small size, artificial nature, low immunogenicity, and many other properties. However, there is no comparative information on the targeting abilities of scaffold polypeptides, which should be taken into account when developing drug delivery systems (DDSs). The present work is the first comprehensive study on the comparison of the effectiveness of different HER2-targeting proteins within the architecture of nanoparticles. Namely, we synthesized trimodal nanoparticles: magnetic, fluorescent, and directed toward HER2 oncomarker on cancer cells. The magnetic particles (MPs) were covalently modified with (i) full-size IgG, 150 kDa, (ii) DARPin_G3, 14 kDa, and (iii) affibody Z_HER2:342, 8 kDa. We showed that the number of DARPin_G3 and affibody Z_HER2:342 molecules conjugated to the nanoparticle surface are 10 and 40 times higher, respectively, than the corresponding value for trastuzumab. Using the methods of magnetic particle quantification (MPQ)-cytometry and confocal microscopy, we showed that all types of the obtained magnetic conjugates specifically labeled HER2-overexpressing cells. Namely, we demonstrated that particle binding to HER2-positive cells is 1113 ± 39 fg/cell for MP*trastuzumab, 1431 ± 186 fg/cell for MP*Z_HER2:342, and 625±21 fg/cell for MP*DARPin_G3, which are 2.77, 2.75, and 2.30 times higher than the corresponding values for control HER2-negative cells. Thus, we showed that the smallest HER2-recognizing polypeptide affibody Z_HER2:342 is more effective in terms of specificity and selectivity in nanoparticle-mediated cell labeling.