Abstract

A murine monoclonal antibody, designated MA-2G9, directed against outer surface protein A (OspA) of the Lyme disease spirochete, Borrelia burgdorferi, has been produced. Antibody MA-2G9, IgG1 subclass, was purified by affinity chromatography on protein G Sepharose column and used for purification of OspA antigen from Borrelia burgdorferi cell lysate. Epitope specificity was studied by Western immunoblotting, using several strains of B. burgdorferi and non-Lyme disease bacteria such as Treponema pallidum and B. hermsii. The MA-2G9 monoclonal antibody reacted specifically with recombinant OspA aas well as with native OspA in sonicated B. burgdorferi strains. No reaction was observed with T. pallidum, Escherichia coli, Staphylococcus aureus and B. hermsii lysates. The MA-2G9 antibody also recognized the denatured form of OspA indicating that it is directed against sequential epitope and not conformational epitope.

References

1
Burgdorfer
W.
Barbour
A.G.
Hayes
S.F.
Benach
J.L.
Grunwaldt
E.
Davis
J.P.
(
1982
)
Lyme disease—a tick-borne spirochetosis?
Science
 ,
216
,
1317
1319
.
2
Steere
A.C.
Grodzicki
R.L.
Kornblatt
A.N.
Craft
J.E.
Barbour
A.G.
Burgdorfer
W.
Schmid
G.P.
Johnson
E.
Malawista
S.E.
(
1983
)
The spirochetal etiology of Lyme disease
N. Engl. J. Med.
 ,
308
,
733
740
.
3
Steere
A.C.
(
1989
)
Lyme disease
N. Engl. J. Med.
 ,
321
,
586
596
.
4
Craft
J.E.
Grodzicki
R.L.
Steere
A.C.
(
1984
)
The antibody response in Lyme disease: evaluation of diagnostic test
J. Infect. Dis.
 ,
149
,
789
795
.
5
Russell
H.
Sampson
J.S.
Schmid
G.P.
Wilkinson
H.W.
Plikaytis
B.
(
1984
)
Enzyme-linked immunosorbent assay and indirect immuno-fluorescence assay for Lyme disease
J. Infect. Dis.
 ,
149
,
465
470
.
6
Magnarelli
L.A.
Anderson
J.F.
Barbour
A.G.
(
1989
)
Enzyme-linked immunosorbent assays for Lyme disease: Reactivity of subunits of Borrelia burgdorferi
J. Infect. Dis.
 ,
159
,
43
49
.
7
Grodzicki
R.L.
Steere
A.C.
(
1988
)
Comparison of immunoblotting and indirect Enzyme-linked immunosorbent assay using different antigen preparations for diagnosing early Lyme disease
J. Infect. Dis.
 ,
157
,
790
797
.
8
Magnarelli
L.A.
Anderson
J.F.
Johnson
R.C.
(
1987
)
Cross-reactivity in serological tests for Lyme disease and other spirochetal infections
J. Infect. Dis.
 ,
156
,
183
187
.
9
Benach
J.L.
Coleman
J.L.
Golightly
M.G.
(
1988
)
A murine IgM monoclonal antibody binds an antigenic determinant in outer surface protein A, an immunodominant basic protein of the Lyme disease spirochete
J. Immunol.
 ,
140
,
265
272
.
10
Hyde
F.W.
Johnson
R.C.
White
T.J.
Shelburne
C.E.
(
1989
)
Detection of antigens in urine of mice and humans infected with B. burgdorferi, etiologic agent of Lyme disease
J. Clin. Microbiol.
 ,
27
,
58
61
.
11
Benach
J.L.
Bosler
E.M.
Hanrahan
J.P.
Coleman
J.L.
Habicht
G.S.
Bast
T.F.
Cameron
D.J.
Ziegler
J.L.
Barbour
A.G.
Burgdorfer
W.
Edelman
R.
Kaslow
R.A.
(
1983
)
Spirochetes isolated from the blood of two patients with Lyme disease
N. Eng. J. Med.
 ,
308
,
740
742
.
12
Simon
M.M.
Schaible
U.E.
Wallich
R.
Kramer
M.D.
(
1991
)
A mouse model for Borrelia burgdorferi infection: approach to a vaccine against Lyme disease
Immunol. Today
 ,
12
,
11
16
.
13
Schaible
U.E.
Kramer
M.D.
Eichmann
K.
Modolell
M.
Museteanu
C.
Simon
M.M.
(
1990
)
Monoclonal antibodies specific for outer surface protein A (OspA) of Borrelia burgdorferi prevent Lyme borreliosis in severe combined immunodeficiency (scid) mice
 
2nd Edn.
,
87
, In
Proc. Natl. Acad. Sci. USA
 , pp
3768
3772
.
14
Dunn
J.J.
Lade
B.N.
Barbour
A.G.
(
1990
)
Outer surface protein A (OspA) from the Lyme disease spirochete, Borrelia burgdorferi: high level expression and purification of a soluble recombinant form of OspA.
Protein Expression and Purification
 ,
1
,
159
168
.
15
Campbell
A.M.
(
1984
)
Monoclonal antibody technology
2nd Edn.
, Vol.
13
, In
Laboratory Techniques in Biochemistry and Molecular Biology
 , pp
120
Elsevier
,
New York
.
16
Laemmli
U.K.
(
1970
)
Cleavage of structural proteins during the assembly of the head of bacteriophage T4
Nature (Lond.)
 ,
227
,
680
685
.
17
Kyhse-Anderson
L.
(
1984
)
Electroblotting of multiple gels: A simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose
J. Biochem. Biophys. Methods
 ,
10
,
203
209
.