Design of acridinium-9-carboxamides and anti-acridinium antibodies for chemiluminescent signal enhancement.
A novel system of signal enhancement is presented in which every labeled antibody is capable of generating a signal. Three chemiluminescent acridinium-9-carboxamide haptens (1, 2, and 3) which incorporated differences in charge and location of the linker were designed and synthesized. Anti-acridinium polyclonal antibodies for each hapten were screened using surface plasmon resonance instrumentation to determine specificity for each hapten. Anti-acridinium 2 antibodies were found to be non-cross-reactive to acridinium 1. This property was exploited to design secondary antibody conjugates which would bind to primary antibodies labeled with 2 yet could still be labeled with the structurally similar acridinium 1. Consequently, both layers contributed to the overall chemiluminescent signal. This format is an advance over other signal amplification formats which employ non-signal-generating, labeled antibodies to construct multilayered systems.
Adamczyk
M
M
Department of Chemistry (9NM), Abbott Diagnostics Division, Abbott Laboratories, Abbott Park, Illinois 60064-6016, USA.
Mattingly
P G
PG
Moore
J A
JA
Pan
Y
Y
Shreder
K
K
Yu
Z
Z
D000166:Acridines / Q000138:chemical synthesis*; D000818:Animals; D000906:Antibodies; D015195:Drug Design; D006241:Haptens / Q000276:immunology*; D007118:Immunoassay / Q000379:methods; D018796:Immunoconjugates / Q000737:chemistry; D008163:Luminescent Measurements; D011817:Rabbits
D000166:Acridines; D000906:Antibodies; D006241:Haptens; D018796:Immunoconjugates; C402305:acridinium-9-carboxamide