PMID- 2420897
OWN - NLM
STAT- MEDLINE
DCOM- 19860514
LR  - 20190723
IS  - 0022-1759 (Print)
IS  - 0022-1759 (Linking)
VI  - 88
IP  - 1
DP  - 1986 Apr 3
TI  - Enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to 
      protein-reactive drugs and metabolites: criteria for identification of antibody 
      activity. Detection and hapten specificity of anti-DNP, anti-captopril and 
      anti-sulphanilamidobenzoic acid.
PG  - 37-44
AB  - Certain hypersensitivity reactions to drugs are thought to depend on coupling of 
      reactive species (the drug itself or a metabolite) to macromolecules, leading to 
      the formation of hapten-carrier conjugates. In assays for the detection of 
      antibodies directed against such reactive species the drug or metabolite must be 
      used in conjugated rather than free form. We describe ELISAs for the detection of 
      anti-dinitrophenyl (DNP), anti-captopril (CP) and anti-sulphanilamidobenzoic acid 
      (SABA) antibodies, in which the wells of microtitre plates are coated with hapten 
      conjugated to protein. We define coating conditions and the following 3 criteria 
      for identification of anti-hapten activity: Immunoglobulin in the test sample 
      binds to the immobilised hapten-protein conjugate, but not to the immobilised 
      protein alone. Binding is inhibited by preincubation of the test sample with 
      protein conjugates incorporating the test hapten, but not by preincubation with 
      the same unconjugated proteins, nor protein conjugates incorporating haptenic 
      groups unrelated to the test hapten. The inhibitory hapten-protein conjugates are 
      shown to be inactive in unrelated antigen-antibody interactions. Binding is 
      blocked by preincubation of the test sample with low molecular weight chemical 
      derivatives of the reactive hapten. The inhibitory derivatives must be shown to 
      be inactive in unrelated antigen-antibody interactions. On the basis of these 
      criteria, IgG anti-DNP and IgG anti-CP were detected in the sera of immunized 
      rabbits. The IgG anti-DNP antibody recognised protein-conjugated DNP, DNP-lysine, 
      N-acetyl-DNP-lysine and DNP-S-glutathione, whereas the IgG anti-CP antibody 
      recognised CP-S-S-protein and CP-S-S-CP. By the same criteria IgG anti-SABA was 
      detected in the sera of immunized mice. The antibody recognised free and 
      protein-conjugated SABA, but not free sulphanilamide.
FAU - Coleman, J W
AU  - Coleman JW
FAU - Yeung, J H
AU  - Yeung JH
FAU - Tingle, M D
AU  - Tingle MD
FAU - Park, B K
AU  - Park BK
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (Antibodies)
RN  - 0 (Azides)
RN  - 0 (Dinitrobenzenes)
RN  - 0 (Epitopes)
RN  - 0 (Haptens)
RN  - 0 (Nitrobenzenes)
RN  - 0 (Proteins)
RN  - 333DO1RDJY (Serotonin)
RN  - 98409-42-8 (3-(2-(4-azidobenzamidino)ethyl)-5-hydroxyindole)
RN  - 9G64RSX1XD (Captopril)
SB  - IM
MH  - Animals
MH  - Antibodies/*analysis
MH  - Antibody Specificity
MH  - Azides/*immunology
MH  - Binding, Competitive
MH  - Captopril/*immunology
MH  - Dinitrobenzenes/*immunology
MH  - Enzyme-Linked Immunosorbent Assay/methods
MH  - Epitopes
MH  - Haptens
MH  - Nitrobenzenes/*immunology
MH  - Proteins/*immunology
MH  - Rabbits
MH  - Serotonin/*analogs & derivatives/immunology
EDAT- 1986/04/03 00:00
MHDA- 1986/04/03 00:01
CRDT- 1986/04/03 00:00
PHST- 1986/04/03 00:00 [pubmed]
PHST- 1986/04/03 00:01 [medline]
PHST- 1986/04/03 00:00 [entrez]
AID - 0022-1759(86)90049-9 [pii]
AID - 10.1016/0022-1759(86)90049-9 [doi]
PST - ppublish
SO  - J Immunol Methods. 1986 Apr 3;88(1):37-44. doi: 10.1016/0022-1759(86)90049-9.