Engineered polymer nanoparticles incorporating l-amino acid groups as affinity reagents for fibrinogen

Yongyan Zhu; Ruixuan Liu; Dengyu Wu; Qianqian Yu; Kenneth J Shea; Quanhong Zhu

doi:10.1016/j.jpha.2020.10.004

Engineered polymer nanoparticles incorporating l-amino acid groups as affinity reagents for fibrinogen

J Pharm Anal. 2021 Oct;11(5):596-602. doi: 10.1016/j.jpha.2020.10.004. Epub 2020 Oct 22.

Authors

Yongyan Zhu^{1

2

3}, Ruixuan Liu¹, Dengyu Wu¹, Qianqian Yu¹, Kenneth J Shea⁴, Quanhong Zhu^{1

2

3}

Affiliations

¹ School of Traditional Chinese Medicine, Southern Medical University, Guangzhou, 510515, China.
² Guangdong Provincial Key Laboratory of Chinese Medicine Pharmaceutics, Guangzhou, 510515, China.
³ Guangdong Provincial Engineering Laboratory of Chinese Medicine Preparation Technology, Guangzhou, 510515, China.
⁴ Department of Chemistry, University of California, Irvine, CA, 92697, USA.

Abstract

Synthetic polymer hydrogel nanoparticles (NPs) were developed to function as abiotic affinity reagents for fibrinogen. These NPs were made using both temperature-sensitive N-isopropyl acrylamide (NIPAm) and l-amino acid monomers. Five kinds of l-amino acids were acryloylated to obtain functional monomers: l-phenylalanine (Phe) and l-leucine (Leu) with hydrophobic side chains, l-glutamic acid (Glu) with negative charges, and l-lysine (Lys) and l-arginine (Arg) with positive charges. After incubating the NPs with fibrinogen, γ-globulin, and human serum albumin (HSA) respectively, the NPs that incorporated N-acryloyl-Arg monomers (AArg@NPs) showed the strongest and most specific binding affinity to fibrinogen, when compared with γ-globulin and HSA. Additionally, the fibrinogen-AArg binding model had the best docking scores, and this may be due to the interaction of positively charged AArg@NPs and the negatively charged fibrinogen D domain and the hydrophobic interaction between them. The specific adsorption of AArg@NPs to fibrinogen was also confirmed by the immunoprecipitation assay, as the AArg@NPs selectively trapped the fibrinogen from a human plasma protein mixture. AArg@NPs had a strong selectivity for, and specificity to, fibrinogen and may be developed as a potential human fibrinogen-specific affinity reagent.

Keywords: Affinity reagent; Amino-acid monomers; Arginine; Fibrinogen; Protein interaction; Synthetic polymer nanoparticles.