Open sandwich ELISA: a novel immunoassay based on the interchain interaction of antibody variable region.
We describe an immunoassay that is based on the interchain interaction of separated VL and VH chains from a single chain antibody variable region. In the presence of antigen, the chains reassociate. VL fragments of anti-hen egg lysozyme (HEL) antibody HyHEL-10 were immobilized on microtiter plates. Samples were coincubated with an M13-displayed VH chain, and assayed with peroxidase-labeled anti-M13 antibody. Signal was detected in direct proportion to the amount of HEL in the sample. Wide dynamic range with < 15 ng/ml sensitivity was attained.
Ueda
H
H
Department of Chemistry and Biotechnology, Graduate School of Engineering, University of Tokyo, Japan. hueda@bio.t.u-tokyo.ac.jp
Tsumoto
K
K
Kubota
K
K
Suzuki
E
E
Nagamune
T
T
Nishimura
H
H
Schueler
P A
PA
Winter
G
G
Kumagai
I
I
Mohoney
W C
WC
D000818:Animals; D000937:Antigen-Antibody Reactions; D001435:Bacteriophages / Q000235:genetics / Q000276:immunology; D015374:Biosensing Techniques; D002645:Chickens; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007128:Immunoglobulin Fragments / Q000235:genetics / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000096:biosynthesis / Q000276:immunology*; D009113:Muramidase / Q000276:immunology*; D009154:Mutation
D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; C055966:immunoglobulin Fv; D009113:Muramidase