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. 2020 May 2;10(5):879.
doi: 10.3390/nano10050879.

XPS Modeling of Immobilized Recombinant Angiogenin and Apoliprotein A1 on Biodegradable Nanofibers

Anton Manakhov  1 , Elizaveta Permyakova  1 , Sergey Ershov  2 , Svetlana Miroshnichenko  1   3 , Mariya Pykhtina  1   3 , Anatoly Beklemishev  1   3 , Andrey Kovalskii  4 , Anastasiya Solovieva  1
Affiliations

XPS Modeling of Immobilized Recombinant Angiogenin and Apoliprotein A1 on Biodegradable Nanofibers

Anton Manakhov et al. Nanomaterials (Basel). .

Abstract

The immobilization of viable proteins is an important step in engineering efficient scaffolds for regenerative medicine. For example, angiogenin, a vascular growth factor, can be considered a neurotrophic factor, influencing the neurogenesis, viability, and migration of neurons. Angiogenin shows an exceptional combination of angiogenic, neurotrophic, neuroprotective, antibacterial, and antioxidant activities. Therefore, this protein is a promising molecule that can be immobilized on carriers used for tissue engineering, particularly for diseases that are complicated by neurotrophic and vascular disorders. Another highly important and viable protein is apoliprotein A1. Nevertheless, the immobilization of these proteins onto promising biodegradable nanofibers has not been tested before. In this work, we carefully studied the immobilization of human recombinant angiogenin and apoliprotein A1 onto plasma-coated nanofibers. We developed a new methodology for the quantification of the protein density of these proteins using X-ray photoelectron spectroscopy (XPS) and modeled the XPS data for angiogenin and apoliprotein A1 (Apo-A1). These findings were also confirmed by the analysis of immobilized Apo-A1 using fluorescent microscopy. The presented methodology was validated by the analysis of fibronectin on the surface of plasma-coated poly(ε-caprolactone) (PCL) nanofibers. This methodology can be expanded for other proteins and it should help to quantify the density of proteins on surfaces using routine XPS data treatment.

Keywords: X-ray photoelectron spectroscopy; angiogenin; biotechnology; nanofibers; plasma; polymers.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
SEM micrographs of PCL-ref (a), PCL-Apo (b), PCL-ANG (c), PCL-COOH (d), PCL-COOH-Apo (e) and PCL-COOH-ANG (f). The size of the bar is 1 µm.
Figure 2
Figure 2
The XPS C1s curve fitting of PCL-ref (a), PCL-Apo (b), PCL-ANG (c), PCL-COOH (d), PCL-COOH-Apo (e), PCL-COOH-ANG (f).
Figure 3
Figure 3
The FT-IR spectra of the PCL nanofibers.
Figure 4
Figure 4
The calculated XPS C1s spectra of apoliprotein A1 (a) and human recombinant angiogenin (b).
Figure 5
Figure 5
A comparison of the XPS C1s spectra: the experimentally measured PCL-Apo (PCL-Apo-Experiment), PCL-ref, and calculated PCL-Apo (PCL-Apo-modeling).
Figure 6
Figure 6
A comparison of the XPS C1s signals for PCL-COOH-Apo, which were measured experimentally and modeled with the different concentrations of protein (x) on the surface. An overview (a) and an enlarged image of the spectrum (b) are given to better show the effect of x on the similarity of the calculated spectrum to the experimental data.
Figure 7
Figure 7
A comparison of the XPS C1s signals of PCL-ANG (a) and PCL-COOH-ANG (b), which were measured experimentally and modeled using Equation (1).
Figure 8
Figure 8
The XPS C1s spectra of the simulated fibronectin (a) and a comparison of the experimentally measured and calculated PCL-COOH-FN (b).
Figure 9
Figure 9
The structure of Fibronectin(FN), human recombinant Angiogenin (hrANG), and Apoliproprotein Apo-A1.
See this image and copyright information in PMC

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