Antibody binding of deletion mutants of Asp f 2, the major Aspergillus fumigatus allergen

Bin Tang; Banani Banerjee; Paul A. Greenberger; Jordan N. Fink; Kevin J. Kelly; Viswanath P. Kurup

doi:10.1006/bbrc.2000.2546

Antibody binding of deletion mutants of Asp f 2, the major Aspergillus fumigatus allergen

Bin Tang^*, Banani Banerjee, Paul A. Greenberger, Jordan N. Fink, Kevin J. Kelly, Viswanath P. Kurup

^*Corresponding author for this work

Medicine, Allergy Division

Research output: Contribution to journal › Article › peer-review

24 Scopus citations

Abstract

Asp f 2, a 268 amino acid major allergen from Aspergillus fumigatus (Af) demonstrated nine linear IgE binding regions. It is not known whether any of these linear epitopes are also conformatory epitopes. Hence, we constructed deletion mutants of Asp f 2 devoid of one or more epitopes, and the IgE binding of these proteins with sera from patients with ABPA was compared with the full-length Asp f 2 expressed in E. coli and Pichia. The Pichia expressed protein reacted weakly with IgE, but strongly with IgG of ABPA sera compared to E. coli expressed Asp f 2. Weak IgE binding only was seen when the C-terminal or N-terminal was deleted, while depletion of both ends negated all reactivity. The monoclonal antibody IL-B8 and IgE and IgG of ABPA sera bound significantly to the Asp f 2 E-4 fragment indicating that the major B-cell epitope is located at the N-terminal end of Asp f 2. (C) 2000 Academic Press.

Original language	English (US)
Pages (from-to)	1128-1135
Number of pages	8
Journal	Biochemical and Biophysical Research Communications
Volume	270
Issue number	3
DOIs	https://doi.org/10.1006/bbrc.2000.2546
State	Published - Apr 21 2000

Funding

This investigation was supported by NIH Grant AI42349 and by Veterans Affairs Medical Research and the Ernest S. Bazley Grant to Northwestern Memorial Hospital and Northwestern University. The technical assistance of Laura Castillo and Nancy Elms and the editorial assistance of Donna Schrubbe are gratefully acknowledged.

Keywords

Asp f 2
Aspergillus fumigatus
Deletion mutants
IgE
Immune response

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

Access to Document

10.1006/bbrc.2000.2546

Cite this

@article{ed379686e5d348149996c2e08cdc6a70,

title = "Antibody binding of deletion mutants of Asp f 2, the major Aspergillus fumigatus allergen",

abstract = "Asp f 2, a 268 amino acid major allergen from Aspergillus fumigatus (Af) demonstrated nine linear IgE binding regions. It is not known whether any of these linear epitopes are also conformatory epitopes. Hence, we constructed deletion mutants of Asp f 2 devoid of one or more epitopes, and the IgE binding of these proteins with sera from patients with ABPA was compared with the full-length Asp f 2 expressed in E. coli and Pichia. The Pichia expressed protein reacted weakly with IgE, but strongly with IgG of ABPA sera compared to E. coli expressed Asp f 2. Weak IgE binding only was seen when the C-terminal or N-terminal was deleted, while depletion of both ends negated all reactivity. The monoclonal antibody IL-B8 and IgE and IgG of ABPA sera bound significantly to the Asp f 2 E-4 fragment indicating that the major B-cell epitope is located at the N-terminal end of Asp f 2. (C) 2000 Academic Press.",

keywords = "Asp f 2, Aspergillus fumigatus, Deletion mutants, IgE, Immune response",

author = "Bin Tang and Banani Banerjee and Greenberger, {Paul A.} and Fink, {Jordan N.} and Kelly, {Kevin J.} and Kurup, {Viswanath P.}",

note = "Funding Information: This investigation was supported by NIH Grant AI42349 and by Veterans Affairs Medical Research and the Ernest S. Bazley Grant to Northwestern Memorial Hospital and Northwestern University. The technical assistance of Laura Castillo and Nancy Elms and the editorial assistance of Donna Schrubbe are gratefully acknowledged.",

year = "2000",

month = apr,

day = "21",

doi = "10.1006/bbrc.2000.2546",

language = "English (US)",

volume = "270",

pages = "1128--1135",

journal = "Biochemical and Biophysical Research Communications",

issn = "0006-291X",

publisher = "Elsevier B.V.",

number = "3",

}

TY - JOUR

T1 - Antibody binding of deletion mutants of Asp f 2, the major Aspergillus fumigatus allergen

AU - Tang, Bin

AU - Banerjee, Banani

AU - Greenberger, Paul A.

AU - Fink, Jordan N.

AU - Kelly, Kevin J.

AU - Kurup, Viswanath P.

N1 - Funding Information: This investigation was supported by NIH Grant AI42349 and by Veterans Affairs Medical Research and the Ernest S. Bazley Grant to Northwestern Memorial Hospital and Northwestern University. The technical assistance of Laura Castillo and Nancy Elms and the editorial assistance of Donna Schrubbe are gratefully acknowledged.

PY - 2000/4/21

Y1 - 2000/4/21

N2 - Asp f 2, a 268 amino acid major allergen from Aspergillus fumigatus (Af) demonstrated nine linear IgE binding regions. It is not known whether any of these linear epitopes are also conformatory epitopes. Hence, we constructed deletion mutants of Asp f 2 devoid of one or more epitopes, and the IgE binding of these proteins with sera from patients with ABPA was compared with the full-length Asp f 2 expressed in E. coli and Pichia. The Pichia expressed protein reacted weakly with IgE, but strongly with IgG of ABPA sera compared to E. coli expressed Asp f 2. Weak IgE binding only was seen when the C-terminal or N-terminal was deleted, while depletion of both ends negated all reactivity. The monoclonal antibody IL-B8 and IgE and IgG of ABPA sera bound significantly to the Asp f 2 E-4 fragment indicating that the major B-cell epitope is located at the N-terminal end of Asp f 2. (C) 2000 Academic Press.

AB - Asp f 2, a 268 amino acid major allergen from Aspergillus fumigatus (Af) demonstrated nine linear IgE binding regions. It is not known whether any of these linear epitopes are also conformatory epitopes. Hence, we constructed deletion mutants of Asp f 2 devoid of one or more epitopes, and the IgE binding of these proteins with sera from patients with ABPA was compared with the full-length Asp f 2 expressed in E. coli and Pichia. The Pichia expressed protein reacted weakly with IgE, but strongly with IgG of ABPA sera compared to E. coli expressed Asp f 2. Weak IgE binding only was seen when the C-terminal or N-terminal was deleted, while depletion of both ends negated all reactivity. The monoclonal antibody IL-B8 and IgE and IgG of ABPA sera bound significantly to the Asp f 2 E-4 fragment indicating that the major B-cell epitope is located at the N-terminal end of Asp f 2. (C) 2000 Academic Press.

KW - Asp f 2

KW - Aspergillus fumigatus

KW - Deletion mutants

KW - IgE

KW - Immune response

UR - http://www.scopus.com/inward/record.url?scp=0034696795&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034696795&partnerID=8YFLogxK

U2 - 10.1006/bbrc.2000.2546

DO - 10.1006/bbrc.2000.2546

M3 - Article

C2 - 10772962

AN - SCOPUS:0034696795

SN - 0006-291X

VL - 270

SP - 1128

EP - 1135

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

IS - 3

ER -

Antibody binding of deletion mutants of Asp f 2, the major Aspergillus fumigatus allergen

Abstract

Funding

Keywords

ASJC Scopus subject areas

Access to Document

Other files and links

Fingerprint

Cite this