Development of a protein nanoparticle platform for targeting EGFR expressing cancer cells

Jakob W. Buecheler, Christopher B. Howard, Christopher J. de Bakker, Stephen Goodall, Martina L. Jones, Thinzar Win, Tao Peng, Cher Heng Tan, Akhil Chopra, Stephen M. Mahler, Sierin Lim

Research output: Contribution to journalArticle

Abstract

BACKGROUND: A range of protein-based nanoparticles has been developed for cancer drug delivery and diagnostics. This includes the E2 protein derived from the pyruvate dehydrogenase complex in Geobacillus stearothermophilus which assembles into a 60-subunit protein cage structure that is capable of encapsulating cancer therapeutics. In this study antibody fragments targeting the epidermal growth factor receptor (EGFR) were tethered to the surface of E2 protein nanoparticles to determine whether the protein nanoparticles could be specifically targeted to EGFR overexpressing cancer cells. RESULTS: Variants of the anti-EGFR antibody fragment and the E2 protein containing specific cysteine residues (E2ΔN17A186C) were conjugated using a maleimide-specific crosslinker. Electron microscopy and dynamic light scattering analysis indicated that the cysteine modified E2 protein correctly assembled into a 25-30nm particle. The conjugation of the anti-EGFR antibody fragment (26 kDa) with a subunit of the E2 protein (26 kDa) was confirmed by mass spectrometry with an estimated molecular weight of 52 kDa. The binding of the conjugated E2 particle to native EGFR on MDA MB 231 cells and recombinant EGFR was confirmed using flow cytometry and biolayer interferometry, respectively. CONCLUSIONS: In this study, proof-of-principle that an EGFR-targeting scFv can be stably conjugated to the cysteine variant E2ΔN17A186C protein nanoparticle without loss of targeting capability has been demonstrated. Conceptually scFv antibody fragments reactive with other important cancer targets could be utilized and presents the opportunity for generation of multi-targeted protein nanoparticles by conjugating various scFvs with different specificities on the same particle.

Original languageEnglish (US)
Pages (from-to)1230-1236
Number of pages7
JournalJournal of Chemical Technology and Biotechnology
Volume90
Issue number7
DOIs
StatePublished - Jul 1 2015
Externally publishedYes

Fingerprint

Epidermal Growth Factor Receptor
Nanoparticles
targeting
cancer
Cells
Immunoglobulin Fragments
Proteins
protein
Neoplasms
Antibodies
Cysteine
antibody
Protein Subunits
Interferometry
Pyruvate Dehydrogenase Complex
Geobacillus stearothermophilus
Single-Chain Antibodies
nanoparticle
Epidermal Growth Factor
Flow cytometry

Keywords

  • Antibody targeting
  • Cancer therapeutic
  • Drug delivery
  • Epidermal growth factor receptor
  • Protein nanoparticle

ASJC Scopus subject areas

  • Chemical Engineering(all)
  • Biotechnology
  • Fuel Technology
  • Renewable Energy, Sustainability and the Environment
  • Pollution
  • Waste Management and Disposal
  • Inorganic Chemistry
  • Organic Chemistry

Cite this

Buecheler, J. W., Howard, C. B., de Bakker, C. J., Goodall, S., Jones, M. L., Win, T., ... Lim, S. (2015). Development of a protein nanoparticle platform for targeting EGFR expressing cancer cells. Journal of Chemical Technology and Biotechnology, 90(7), 1230-1236. https://doi.org/10.1002/jctb.4545

Development of a protein nanoparticle platform for targeting EGFR expressing cancer cells. / Buecheler, Jakob W.; Howard, Christopher B.; de Bakker, Christopher J.; Goodall, Stephen; Jones, Martina L.; Win, Thinzar; Peng, Tao; Tan, Cher Heng; Chopra, Akhil; Mahler, Stephen M.; Lim, Sierin.

In: Journal of Chemical Technology and Biotechnology, Vol. 90, No. 7, 01.07.2015, p. 1230-1236.

Research output: Contribution to journalArticle

Buecheler, JW, Howard, CB, de Bakker, CJ, Goodall, S, Jones, ML, Win, T, Peng, T, Tan, CH, Chopra, A, Mahler, SM & Lim, S 2015, 'Development of a protein nanoparticle platform for targeting EGFR expressing cancer cells', Journal of Chemical Technology and Biotechnology, vol. 90, no. 7, pp. 1230-1236. https://doi.org/10.1002/jctb.4545
Buecheler, Jakob W. ; Howard, Christopher B. ; de Bakker, Christopher J. ; Goodall, Stephen ; Jones, Martina L. ; Win, Thinzar ; Peng, Tao ; Tan, Cher Heng ; Chopra, Akhil ; Mahler, Stephen M. ; Lim, Sierin. / Development of a protein nanoparticle platform for targeting EGFR expressing cancer cells. In: Journal of Chemical Technology and Biotechnology. 2015 ; Vol. 90, No. 7. pp. 1230-1236.
@article{ce46215ef9f54898b6764cb8fccb9b11,
title = "Development of a protein nanoparticle platform for targeting EGFR expressing cancer cells",
abstract = "BACKGROUND: A range of protein-based nanoparticles has been developed for cancer drug delivery and diagnostics. This includes the E2 protein derived from the pyruvate dehydrogenase complex in Geobacillus stearothermophilus which assembles into a 60-subunit protein cage structure that is capable of encapsulating cancer therapeutics. In this study antibody fragments targeting the epidermal growth factor receptor (EGFR) were tethered to the surface of E2 protein nanoparticles to determine whether the protein nanoparticles could be specifically targeted to EGFR overexpressing cancer cells. RESULTS: Variants of the anti-EGFR antibody fragment and the E2 protein containing specific cysteine residues (E2ΔN17A186C) were conjugated using a maleimide-specific crosslinker. Electron microscopy and dynamic light scattering analysis indicated that the cysteine modified E2 protein correctly assembled into a 25-30nm particle. The conjugation of the anti-EGFR antibody fragment (26 kDa) with a subunit of the E2 protein (26 kDa) was confirmed by mass spectrometry with an estimated molecular weight of 52 kDa. The binding of the conjugated E2 particle to native EGFR on MDA MB 231 cells and recombinant EGFR was confirmed using flow cytometry and biolayer interferometry, respectively. CONCLUSIONS: In this study, proof-of-principle that an EGFR-targeting scFv can be stably conjugated to the cysteine variant E2ΔN17A186C protein nanoparticle without loss of targeting capability has been demonstrated. Conceptually scFv antibody fragments reactive with other important cancer targets could be utilized and presents the opportunity for generation of multi-targeted protein nanoparticles by conjugating various scFvs with different specificities on the same particle.",
keywords = "Antibody targeting, Cancer therapeutic, Drug delivery, Epidermal growth factor receptor, Protein nanoparticle",
author = "Buecheler, {Jakob W.} and Howard, {Christopher B.} and {de Bakker}, {Christopher J.} and Stephen Goodall and Jones, {Martina L.} and Thinzar Win and Tao Peng and Tan, {Cher Heng} and Akhil Chopra and Mahler, {Stephen M.} and Sierin Lim",
year = "2015",
month = "7",
day = "1",
doi = "10.1002/jctb.4545",
language = "English (US)",
volume = "90",
pages = "1230--1236",
journal = "Journal of Chemical Technology and Biotechnology",
issn = "0268-2575",
publisher = "John Wiley and Sons Ltd",
number = "7",

}

TY - JOUR

T1 - Development of a protein nanoparticle platform for targeting EGFR expressing cancer cells

AU - Buecheler, Jakob W.

AU - Howard, Christopher B.

AU - de Bakker, Christopher J.

AU - Goodall, Stephen

AU - Jones, Martina L.

AU - Win, Thinzar

AU - Peng, Tao

AU - Tan, Cher Heng

AU - Chopra, Akhil

AU - Mahler, Stephen M.

AU - Lim, Sierin

PY - 2015/7/1

Y1 - 2015/7/1

N2 - BACKGROUND: A range of protein-based nanoparticles has been developed for cancer drug delivery and diagnostics. This includes the E2 protein derived from the pyruvate dehydrogenase complex in Geobacillus stearothermophilus which assembles into a 60-subunit protein cage structure that is capable of encapsulating cancer therapeutics. In this study antibody fragments targeting the epidermal growth factor receptor (EGFR) were tethered to the surface of E2 protein nanoparticles to determine whether the protein nanoparticles could be specifically targeted to EGFR overexpressing cancer cells. RESULTS: Variants of the anti-EGFR antibody fragment and the E2 protein containing specific cysteine residues (E2ΔN17A186C) were conjugated using a maleimide-specific crosslinker. Electron microscopy and dynamic light scattering analysis indicated that the cysteine modified E2 protein correctly assembled into a 25-30nm particle. The conjugation of the anti-EGFR antibody fragment (26 kDa) with a subunit of the E2 protein (26 kDa) was confirmed by mass spectrometry with an estimated molecular weight of 52 kDa. The binding of the conjugated E2 particle to native EGFR on MDA MB 231 cells and recombinant EGFR was confirmed using flow cytometry and biolayer interferometry, respectively. CONCLUSIONS: In this study, proof-of-principle that an EGFR-targeting scFv can be stably conjugated to the cysteine variant E2ΔN17A186C protein nanoparticle without loss of targeting capability has been demonstrated. Conceptually scFv antibody fragments reactive with other important cancer targets could be utilized and presents the opportunity for generation of multi-targeted protein nanoparticles by conjugating various scFvs with different specificities on the same particle.

AB - BACKGROUND: A range of protein-based nanoparticles has been developed for cancer drug delivery and diagnostics. This includes the E2 protein derived from the pyruvate dehydrogenase complex in Geobacillus stearothermophilus which assembles into a 60-subunit protein cage structure that is capable of encapsulating cancer therapeutics. In this study antibody fragments targeting the epidermal growth factor receptor (EGFR) were tethered to the surface of E2 protein nanoparticles to determine whether the protein nanoparticles could be specifically targeted to EGFR overexpressing cancer cells. RESULTS: Variants of the anti-EGFR antibody fragment and the E2 protein containing specific cysteine residues (E2ΔN17A186C) were conjugated using a maleimide-specific crosslinker. Electron microscopy and dynamic light scattering analysis indicated that the cysteine modified E2 protein correctly assembled into a 25-30nm particle. The conjugation of the anti-EGFR antibody fragment (26 kDa) with a subunit of the E2 protein (26 kDa) was confirmed by mass spectrometry with an estimated molecular weight of 52 kDa. The binding of the conjugated E2 particle to native EGFR on MDA MB 231 cells and recombinant EGFR was confirmed using flow cytometry and biolayer interferometry, respectively. CONCLUSIONS: In this study, proof-of-principle that an EGFR-targeting scFv can be stably conjugated to the cysteine variant E2ΔN17A186C protein nanoparticle without loss of targeting capability has been demonstrated. Conceptually scFv antibody fragments reactive with other important cancer targets could be utilized and presents the opportunity for generation of multi-targeted protein nanoparticles by conjugating various scFvs with different specificities on the same particle.

KW - Antibody targeting

KW - Cancer therapeutic

KW - Drug delivery

KW - Epidermal growth factor receptor

KW - Protein nanoparticle

UR - http://www.scopus.com/inward/record.url?scp=84930335371&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84930335371&partnerID=8YFLogxK

U2 - 10.1002/jctb.4545

DO - 10.1002/jctb.4545

M3 - Article

AN - SCOPUS:84930335371

VL - 90

SP - 1230

EP - 1236

JO - Journal of Chemical Technology and Biotechnology

JF - Journal of Chemical Technology and Biotechnology

SN - 0268-2575

IS - 7

ER -