Flow imaging microscopy as a novel tool for high-throughput evaluation of elastin-like polymer coacervates

PLoS One. 2019 May 9;14(5):e0216406. doi: 10.1371/journal.pone.0216406. eCollection 2019.

Abstract

Biological and bioinspired polymer microparticles have broad biomedical and industrial applications, including drug delivery, tissue engineering, surface modification, environmental remediation, imaging, and sensing. Full realization of the potential of biopolymer microparticles will require methods for rigorous characterization of particle sizes, morphologies, and dynamics, so that researchers may correlate particle characteristics with synthesis methods and desired functions. Toward this end, we evaluated biopolymer microparticles using flow imaging microscopy. This technology is widely used in the biopharmaceutical industry but is not yet well-known among the materials community. Our polymer, a genetically engineered elastin-like polypeptide (ELP), self-assembles into micron-scale coacervates. We performed flow imaging of ELP coacervates using two different instruments, one with a lower size limit of approximately 2 microns, the other with a lower size limit of approximately 300 nanometers. We validated flow imaging results by comparison with dynamic light scattering and atomic force microscopy analyses. We explored the effects of various solvent conditions on ELP coacervate size, morphology, and behavior, such as the dispersion of single particles versus aggregates. We found that flow imaging is a superior tool for rapid and thorough particle analysis of ELP coacervates in solution. We anticipate that researchers studying many types of microscale protein or polymer assemblies will be interested in flow imaging as a tool for quantitative, solution-based characterization.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Drug Delivery Systems*
  • Drug Evaluation, Preclinical
  • Elastin / chemistry*
  • Microscopy*

Substances

  • Elastin

Associated data

  • Dryad/10.5061/dryad.14hq8cd

Grants and funding

Grants from the National Science Foundation (https://www.nsf.gov) to ERMB (NSF 1638893) and JMH (NSF 1638896) were used to support personnel and supply costs. ERMB is a co-PI on NSF MRI grants 1531298 and 1624984, which were used for the acquisition of the Cypher ES AFM and FlowCam VS instruments, respectively. LM and WP were supported by Summer Undergraduate Research Experience fellowships provided by the University of New England’s College of Arts and Sciences. LM was also supported by a NASA/Maine Space Grant Consortium award. NG is employed at Fluid Imaging Technologies, Inc., makers of the FlowCam. Fluid Imaging Technologies, Inc. provided support in the form of salary for author NG and material resources in the form of access to their FlowCam Nano instrument. The specific roles of the authors are articulated in the Author Contribution section. The funders had no additional roles in study design, data collection and analysis, decision to publish, or preparation of the manuscript. There was no additional external funding received for this study.