Phthalic anhydride allergy: development and characterization of optimized hapten-carrier conjugates for improved diagnosis.
BACKGROUND
At present the diagnosis of IgE-mediated hypersensitivity to phthalic anhydride (PA) is based on conjugates that are not characterized or standardized. The aim of this study was to develop optimized and molecularly characterized PA conjugates that can be used to improve the diagnosis of PA-allergy.


METHODS
The PA conjugates were synthesized and the number of haptens bound on a carrier protein was estimated by matrix-assisted laser desorption/ionization time of light (MALDI-TOF) mass spectrometry. The ability of conjugates to bind IgE and IgG antibodies was measured by enzyme-linked immunosorbent assay (ELISA). Reactivity of the conjugates in vivo was evaluated by skin prick testing.


RESULTS
The most active IgE-binding conjugates had a PA : HSA molar ratio of 80 : 1. In the optimal conjugates the average numbers of PA haptens per carrier molecule of human serum albumin (HSA) were 14-16. In ELISA, all 13 patients and none of the 20 controls had IgE antibodies to optimized PA conjugate. The sensitivity and specificity of the ELISA was comparable to commercial CAP RAST. PA conjugates elicited positive test results in skin prick testing showing that conjugates are immunologically active also in vivo.


CONCLUSIONS
These results indicate that optimized and molecularly characterized PA-HSA conjugates can be used both in vitro and in vivo assays to improve the diagnosis of PA allergy.
Pakarinen
M
M
Department of Industrial Hygiene and Toxicology, Finnish Institute of Occupational Health, Helsinki, Finland.
Koivuluhta
M
M
Kalkkinen
N
N
Keskinen
H
H
Nordman
H
H
Estlander
T
T
Tupasela
O
O
Jolanki
R
R
Lauerma
A I
AI
Pfäffli
P
P
Alenius
H
H
D000328:Adult; D001667:Binding, Competitive / Q000276:immunology; D002352:Carrier Proteins / Q000032:analysis / Q000276:immunology; D004342:Drug Hypersensitivity / Q000175:diagnosis / Q000209:etiology / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006241:Haptens / Q000145:classification / Q000276:immunology; D006801:Humans; D006969:Hypersensitivity, Immediate / Q000139:chemically induced; D007073:Immunoglobulin E / Q000032:analysis / Q000276:immunology; D007074:Immunoglobulin G / Q000032:analysis / Q000276:immunology; D008297:Male; D008875:Middle Aged; D010796:Phthalic Anhydrides / Q000009:adverse effects* / Q000032:analysis / Q000276:immunology*; D011852:Radioallergosorbent Test; D012709:Serum Albumin / Q000145:classification / Q000276:immunology; D012882:Skin Tests
D002352:Carrier Proteins; D006241:Haptens; D007074:Immunoglobulin G; D010796:Phthalic Anhydrides; D012709:Serum Albumin; D007073:Immunoglobulin E; C043103:phthalic anhydride