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Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...

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Biomolecular Imaging of Cellular Uptake of Nanoparticles using Multimodal Nonlinear Optical Microscopy
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Kinetic quantification of protein polymer nanoparticles using non-invasive imaging.

S M Janib1, S Liu, R Park

  • 1Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA 90033-9121, USA.

Integrative Biology : Quantitative Biosciences From Nano to Macro
|October 25, 2012
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Protein polymers form nanoparticles for cancer nanomedicine. Positron emission tomography (PET) imaging tracked these elastin-like polypeptide (ELP) nanoparticles, revealing molecular weight, not nanostructure, dictates their pharmacokinetics and tumor accumulation.

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Area of Science:

  • Biomaterials Science
  • Nanotechnology
  • Molecular Imaging

Background:

  • Protein polymers, specifically elastin-like polypeptides (ELPs), self-assemble into nanoparticles with potential as cancer nanomedicines.
  • Positron emission tomography (PET) offers sensitive and quantitative molecular imaging crucial for tracking nanotherapeutics.

Purpose of the Study:

  • To investigate the in vivo tracking of ELP-based protein polymer nanoparticles using microPET imaging over several days.
  • To develop and apply a pharmacokinetic model to analyze nanoparticle distribution and behavior.

Main Methods:

  • Site-specific conjugation of polypeptides with AmBaSar chelator for complexation with Copper-64 ((64)Cu).
  • MicroPET imaging in an orthotopic breast cancer model to track four different ELP-based protein polymers.
  • Development of a six-compartment image-driven pharmacokinetic model for data analysis.

Main Results:

  • Nanoparticle formation minimally affected pharmacokinetics or tumor accumulation compared to free ELPs of similar molecular weight.
  • ELP molecular weight was the primary determinant of polymer fate, with longer ELPs exhibiting significantly longer circulation half-lives.
  • Long ELPs (74 kD) had an 8.7-hour half-life, while short ELPs (37 kD) had a 2.1-hour half-life.

Conclusions:

  • ELP molecular weight, rather than nanostructure, is the key factor governing the pharmacokinetic behavior of these protein polymer nanoparticles.
  • ELP-based protein polymers represent a promising platform for developing theranostic nanoparticles that can be monitored with clinical PET scanners.