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Specific Antisera Against the Catecholamines: L‐3,4‐Dihydroxyphenylalanine, Dopamine, Noradrenaline, and Octopamine Tested by an Enzyme‐Linked Immunosorbent Assay
Author(s) -
Mons Nicole,
Geffard Michel
Publication year - 1987
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1987.tb05743.x
Subject(s) - immunogen , antiserum , octopamine (neurotransmitter) , chemistry , biochemistry , bovine serum albumin , dopamine , enzyme , tyramine , serum albumin , biogenic amine , catechol , antibody , microbiology and biotechnology , biology , monoclonal antibody , neurotransmitter , endocrinology , serotonin , immunology , receptor
Antisera were raised against L‐3,4‐dihydroxyphe‐nylalanine (L‐DOPA), dopamine (DA), noradrenaline (NA), and octopamine (OA). This was achieved by coupling each molecule to bovine serum albumin or human serum albumin using glutaraldehyde. The conjugated aromatic amines were kept in a reducing medium containing sodium metabisulfite. Antiserum specificity was tested using an enzyme‐linked immunosorbent assay method for catecholamines. Competition experiments were done between the immunogen coated on the well plates and each catechol‐amine, either in the free state or in conjugated form, previously incubated with an antiserum. In each case, the non‐conjugated compound was poorly recognized. The nonre‐duced conjugates of L‐DOPA and DA were well recognized, whereas those of NA and OA were poorly immunoreactive. The cross‐reactivity ratios established in the competition experiments allowed the specificity of the immune response to be defined. In each case, it was found to be high. The results suggest that the antibodies of L‐DOPA and DA antisera recognize preferentially the catechol moiety, whereas for the anti‐NA and anti‐OA antibodies, the lateral chain is important.