Radioimmunoassay for albuterol using a monoclonal antibody: application for direct quantification in horse urine.
A monoclonal antibody was synthesized in mouse against the O-(3-carboxypropionyl) derivative of albuterol linked to bovine serum albumin. Isotyping of this material revealed the IgG1 class characterized by an affinity constant of 1.03 nM-1 and a density of sites of 0.55 nM. This antibody was found specific as its cross-reactivity to structurally related molecules was less than 1% except for clenbuterol (75%). A radioimmunoassay was set up with culture supernatant (final dilution 1/1000) and [3H] albuterol. The calibration curve was characterized by a maximum binding of 28%, an ED50 of 1.15 pmol per tube, the detection limit was 28.8 fmol/tube and the linearity of the response was up to 39.8 pmol/tube. This RIA method has been used for direct quantitation of albuterol in horse urine without any clean-up or extraction step.
Adam
A
A
Faculté de pharmacie, Université de Montréal, Québec, Canada.
Ong
H
H
Sondag
D
D
Rapaille
A
A
Marleau
S
S
Bellemare
M
M
Raymond
P
P
Giroux
D
D
Loo
J K
JK
Beaulieu
N
N
D000420:Albuterol / Q000652:urine*; D000818:Animals; D000911:Antibodies, Monoclonal; D000915:Antibody Affinity; D000918:Antibody Specificity; D005260:Female; D006736:Horses / Q000652:urine*; D007132:Immunoglobulin Isotypes; D051379:Mice; D011863:Radioimmunoassay; D015203:Reproducibility of Results
D000911:Antibodies, Monoclonal; D007132:Immunoglobulin Isotypes; D000420:Albuterol