Determination of retinoid activity by an enzyme-linked immunosorbent assay.
In normal human keratinocytes, retinoic acid suppresses the expression of the plasma membrane associated enzyme transglutaminase (TGm) at the pretranslational level. This finding led us to develop an enzyme-linked immunosorbent assay (ELISA) for the evaluation of the biological activity of retinoids, i.e., natural and synthetic derivatives of vitamin A. In this assay, keratinocytes are cultured in a 96-well cluster in the presence of different retinoid concentrations. The expression of TGm is then quantified, without any extraction or purification step, using a TGm-specific monoclonal antibody and a peroxidase-conjugated secondary antibody. The dose-response curves obtained show this ELISA to be a sensitive and reproducible assay to determine the potency of retinoids.
Centre International de Recherches Dermatologiques GALDERMA (CIRD GALDERMA), Sophia Antipolis, Valbonne, France.
S
S
Michel
A
A
Courseaux
C
C
Miquel
J M
JM
Bernardon
R
R
Schmidt
B
B
Shroot
S M
SM
Thacher
U
U
Reichert
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002138:Calibration; D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D005069:Evaluation Studies as Topic; D005404:Fixatives; D006801:Humans; D051379:Mice; D012176:Retinoids / Q000378:metabolism*; D012680:Sensitivity and Specificity; D011503:Transglutaminases / Q000276:immunology / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D005404:Fixatives; D012176:Retinoids; D011503:Transglutaminases