Conformational analysis of hepatitis B surface antigen fusions in an Agrobacterium-mediated transient expression system

Zhong Huang, Hugh Mason

Research output: Contribution to journalArticle

53 Citations (Scopus)

Abstract

Vaccine antigens have been successfully produced in transgenic plants for oral immunization. Recently, a fusion strategy has been adopted to produce multicomponent vaccines and to target antigens to mucosal sites for enhanced oral immunogenicity. However, antigen fusions may not be folded correctly due to steric hindrance and may thus lose their potency. Here, we describe an Agrobacterium-mediated transient assay that provides enough antigen-expressing material at 2 days post-transfection to evaluate antigen conformation. Using the hepatitis B surface antigen (HBsAg) as a model antigen and the green fluorescent protein (GFP) as a model fusion partner, we showed that transiently expressed HBsAg and an HBsAg fusion with GFP at the N-terminus (GFP:HBsAg), but not the HBsAg fusion with GFP at the C-terminus (HBsAg:GFP), formed the 'a' determinant and virus-like particles (VLPs), similar to yeast-derived vaccine HBsAg. Thus, it is feasible to modify the HBsAg with an N-terminal fusion of up to 239 amino acids without altering its major antigenic properties. Our results also demonstrate that the Agrobacterium-mediated transient expression system can be used to evaluate the conformation of plant-based vaccines or other pharmaceutical proteins in a high-throughput manner.

Original languageEnglish (US)
Pages (from-to)241-249
Number of pages9
JournalPlant Biotechnology Journal
Volume2
Issue number3
DOIs
StatePublished - May 2004

Fingerprint

Agrobacterium
Hepatitis B Surface Antigens
Green Fluorescent Proteins
green fluorescent protein
antigens
Antigens
Vaccines
vaccines
oral vaccination
biopharmaceuticals
virus-like particles
hepatitis B antigens
Genetically Modified Plants
transfection
Protein C
Virion
Transfection
transgenic plants
mouth
Immunization

Keywords

  • Fusion
  • Green fluorescent protein
  • Hepatitis B surface antigen
  • Transient expression
  • Vaccine
  • Virus-like particle

ASJC Scopus subject areas

  • Plant Science

Cite this

Conformational analysis of hepatitis B surface antigen fusions in an Agrobacterium-mediated transient expression system. / Huang, Zhong; Mason, Hugh.

In: Plant Biotechnology Journal, Vol. 2, No. 3, 05.2004, p. 241-249.

Research output: Contribution to journalArticle

@article{162018ac69f646c8aea4fe683da30be9,
title = "Conformational analysis of hepatitis B surface antigen fusions in an Agrobacterium-mediated transient expression system",
abstract = "Vaccine antigens have been successfully produced in transgenic plants for oral immunization. Recently, a fusion strategy has been adopted to produce multicomponent vaccines and to target antigens to mucosal sites for enhanced oral immunogenicity. However, antigen fusions may not be folded correctly due to steric hindrance and may thus lose their potency. Here, we describe an Agrobacterium-mediated transient assay that provides enough antigen-expressing material at 2 days post-transfection to evaluate antigen conformation. Using the hepatitis B surface antigen (HBsAg) as a model antigen and the green fluorescent protein (GFP) as a model fusion partner, we showed that transiently expressed HBsAg and an HBsAg fusion with GFP at the N-terminus (GFP:HBsAg), but not the HBsAg fusion with GFP at the C-terminus (HBsAg:GFP), formed the 'a' determinant and virus-like particles (VLPs), similar to yeast-derived vaccine HBsAg. Thus, it is feasible to modify the HBsAg with an N-terminal fusion of up to 239 amino acids without altering its major antigenic properties. Our results also demonstrate that the Agrobacterium-mediated transient expression system can be used to evaluate the conformation of plant-based vaccines or other pharmaceutical proteins in a high-throughput manner.",
keywords = "Fusion, Green fluorescent protein, Hepatitis B surface antigen, Transient expression, Vaccine, Virus-like particle",
author = "Zhong Huang and Hugh Mason",
year = "2004",
month = "5",
doi = "10.1111/j.1467-7652.2004.00068.x",
language = "English (US)",
volume = "2",
pages = "241--249",
journal = "Plant Biotechnology Journal",
issn = "1467-7644",
publisher = "Wiley-Blackwell",
number = "3",

}

TY - JOUR

T1 - Conformational analysis of hepatitis B surface antigen fusions in an Agrobacterium-mediated transient expression system

AU - Huang, Zhong

AU - Mason, Hugh

PY - 2004/5

Y1 - 2004/5

N2 - Vaccine antigens have been successfully produced in transgenic plants for oral immunization. Recently, a fusion strategy has been adopted to produce multicomponent vaccines and to target antigens to mucosal sites for enhanced oral immunogenicity. However, antigen fusions may not be folded correctly due to steric hindrance and may thus lose their potency. Here, we describe an Agrobacterium-mediated transient assay that provides enough antigen-expressing material at 2 days post-transfection to evaluate antigen conformation. Using the hepatitis B surface antigen (HBsAg) as a model antigen and the green fluorescent protein (GFP) as a model fusion partner, we showed that transiently expressed HBsAg and an HBsAg fusion with GFP at the N-terminus (GFP:HBsAg), but not the HBsAg fusion with GFP at the C-terminus (HBsAg:GFP), formed the 'a' determinant and virus-like particles (VLPs), similar to yeast-derived vaccine HBsAg. Thus, it is feasible to modify the HBsAg with an N-terminal fusion of up to 239 amino acids without altering its major antigenic properties. Our results also demonstrate that the Agrobacterium-mediated transient expression system can be used to evaluate the conformation of plant-based vaccines or other pharmaceutical proteins in a high-throughput manner.

AB - Vaccine antigens have been successfully produced in transgenic plants for oral immunization. Recently, a fusion strategy has been adopted to produce multicomponent vaccines and to target antigens to mucosal sites for enhanced oral immunogenicity. However, antigen fusions may not be folded correctly due to steric hindrance and may thus lose their potency. Here, we describe an Agrobacterium-mediated transient assay that provides enough antigen-expressing material at 2 days post-transfection to evaluate antigen conformation. Using the hepatitis B surface antigen (HBsAg) as a model antigen and the green fluorescent protein (GFP) as a model fusion partner, we showed that transiently expressed HBsAg and an HBsAg fusion with GFP at the N-terminus (GFP:HBsAg), but not the HBsAg fusion with GFP at the C-terminus (HBsAg:GFP), formed the 'a' determinant and virus-like particles (VLPs), similar to yeast-derived vaccine HBsAg. Thus, it is feasible to modify the HBsAg with an N-terminal fusion of up to 239 amino acids without altering its major antigenic properties. Our results also demonstrate that the Agrobacterium-mediated transient expression system can be used to evaluate the conformation of plant-based vaccines or other pharmaceutical proteins in a high-throughput manner.

KW - Fusion

KW - Green fluorescent protein

KW - Hepatitis B surface antigen

KW - Transient expression

KW - Vaccine

KW - Virus-like particle

UR - http://www.scopus.com/inward/record.url?scp=13244262550&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=13244262550&partnerID=8YFLogxK

U2 - 10.1111/j.1467-7652.2004.00068.x

DO - 10.1111/j.1467-7652.2004.00068.x

M3 - Article

VL - 2

SP - 241

EP - 249

JO - Plant Biotechnology Journal

JF - Plant Biotechnology Journal

SN - 1467-7644

IS - 3

ER -