Assessing sequence plasticity of a virus-like nanoparticle by evolution toward a versatile scaffold for vaccines and drug delivery.

نویسندگان

  • Yuan Lu
  • Wei Chan
  • Benjamin Y Ko
  • Christopher C VanLang
  • James R Swartz
چکیده

Virus-like particles (VLPs) have been extensively explored as nanoparticle vehicles for many applications in biotechnology (e.g., vaccines, drug delivery, imaging agents, biocatalysts). However, amino acid sequence plasticity relative to subunit expression and nanoparticle assembly has not been explored. Whereas the hepatitis B core protein (HBc) VLP appears to be the most promising model for fundamental and applied studies; particle instability, antigen fusion limitations, and intrinsic immunogenicity have limited its development. Here, we apply Escherichia coli-based cell-free protein synthesis (CFPS) to rapidly produce and screen HBc protein variants that still self-assemble into VLPs. To improve nanoparticle stability, artificial covalent disulfide bridges were introduced throughout the VLP. Negative charges on the HBc VLP surface were then reduced to improve surface conjugation. However, removal of surface negative charges caused low subunit solubility and poor VLP assembly. Solubility and assembly as well as surface conjugation were greatly improved by transplanting a rare spike region onto the common shell structure. The newly stabilized and extensively modified HBc VLP had almost no immunogenicity in mice, demonstrating great promise for medical applications. This study introduces a general paradigm for functional improvement of complex protein assemblies such as VLPs. This is the first study, to our knowledge, to systematically explore the sequence plasticity of viral capsids as an approach to defining structure function relationships for viral capsid proteins. Our observations on the unexpected importance of the HBc spike tip charged state may also suggest new mechanistic routes toward viral therapeutics that block capsid assembly.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Truncated Hepatitis B virus like nanoparticles: A novel drug delivery platform for cancer therapy

Nowadays, Nano-sized drug delivery systems have been studied extensively for theirpotential in cancer therapy. Various drug nanocarriers are being developed including liposomes, micelles, and Virus like nanoparticles (VLNPs). VLNPs offer many advantages for developing smart drug delivery systems due to their precise and repeated structures and relatively large cargo capacities. Truncated ...

متن کامل

Re-design of Downstream Processing Techniques for Nanoparticulate Bioproducts

There has been much interest generated in the recovery of nanoparticulate (nanoparticle) bioproducts(Second generation of biotechnological products) such as plasmid DNA and viruses as putative gene therapyvectors, macromolecular assemblies as drug delivery vehicles and virus-like particles as vaccine components.Such product must be manufactured in advanced stages of purity, ma...

متن کامل

Imatinib loaded pegylated Poly Propylene Imine dendrimer for delivery to leukemic cells; fabrication of formulation and evaluation

PEGylated polypropyleneimine (PPI) dendritic scaffold was used for the delivery of an anti-leukemic drug, Imatinib. The current study evolves and emerges the use PEGylated PPI dendritic scaffold for the delivery of this drug. In this Imatinib was synthesized and loaded with PEGylated PPI dendritic scaffold. Parameters such as FT-IR, NMR, SEM, drug release, DSC and hemolytictoxicity are required...

متن کامل

Imatinib loaded pegylated Poly Propylene Imine dendrimer for delivery to leukemic cells; fabrication of formulation and evaluation

PEGylated polypropyleneimine (PPI) dendritic scaffold was used for the delivery of an anti-leukemic drug, Imatinib. The current study evolves and emerges the use PEGylated PPI dendritic scaffold for the delivery of this drug. In this Imatinib was synthesized and loaded with PEGylated PPI dendritic scaffold. Parameters such as FT-IR, NMR, SEM, drug release, DSC and hemolytictoxicity are required...

متن کامل

Au Nanoparticle Loaded with 6-Thioguanine Anticancer Drug as a New Strategy for Drug Delivery

In this study we suggested a new strategy for drug delivery of 6-thioguanine (6-TG) as a cancer drug by loading of this thiolic drug at a surface of Au nanoparticles. For this goal, we synthesized Au nanoparticle (Au/NPs) by reduction of tetrachloroauric (III) acid solutions by sodium borohydride and characterized Au/NPs by X-ray powder diffraction (XRD), dynamic light scattering (DLS), Ultravi...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:
  • Proceedings of the National Academy of Sciences of the United States of America

دوره 112 40  شماره 

صفحات  -

تاریخ انتشار 2015