Rabbit antibody detection with RNA aptamers.
Antibody-based detection systems are widely used, but in the cases of immunoprecipitations and enzyme-linked immunoassays, they can be laborious. These techniques require the preparation of at least two kinds of non-cross-reactive immunoglobulin Gs (IgGs), usually made from different species against the single target molecule. Aptamers composed of nucleic acids possess strict recognition ability for the target molecule's three-dimensional structure and, thus, are considered to act like IgG. In this study, experimental trials were designed to combine the advantages of IgG and aptamers. For this purpose, aptamers against rabbit IgG were identified by in vitro selection. One of the obtained aptamers had a dissociation constant lower than 15 pM to the rabbit IgG. It bound specifically to the constant region of the rabbit IgG, and no binding was observed with mouse or goat IgG. Moreover, this aptamer recognized only the native form of rabbit IgG and could not bind the sodium dodecyl sulfate (SDS)-denatured form. These features show the advantage of using the aptamer as a secondary probing agent rather than the usual secondary antibodies.
Yoshida
Yoshihito
Y
VALWAY Technology Center, NEC Soft, Ltd., Tokyo 136-8627, Japan.
Sakai
Nobuya
N
Masuda
Hiromi
H
Furuichi
Makio
M
Nishikawa
Fumiko
F
Nishikawa
Satoshi
S
Mizuno
Hiroshi
H
Waga
Iwao
I
D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000276:immunology / Q000378:metabolism*; D052157:Aptamers, Nucleotide / Q000032:analysis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D015153:Blotting, Western / Q000379:methods*; D003429:Cross Reactions; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D009691:Nucleic Acid Denaturation; D011817:Rabbits; D013379:Substrate Specificity
D000906:Antibodies; D052157:Aptamers, Nucleotide; D007074:Immunoglobulin G