PMID- 6851184
OWN - NLM
STAT- MEDLINE
DCOM- 19830708
LR  - 20190706
IS  - 0009-8981 (Print)
IS  - 0009-8981 (Linking)
VI  - 130
IP  - 1
DP  - 1983 May 9
TI  - The development of a rapid ELISA for IgE utilizing commercially available 
      reagents.
PG  - 71-83
AB  - An enzyme-linked immunosorbent assay (ELISA) for human immunoglobin E (IgE) has 
      been developed. To coat the polystyrene tubes (the solid phase) an antibody 
      concentration of 6 mg/l in sodium carbonate buffer, pH 9.8, at 37 degrees C for 
      24 h was found superior to other conditions. The maximum amount of globulin 
      adsorbed to the polystyrene surface was estimated to be 2.7 mg X m-2. This is 
      consistent with a monolayer being adsorbed. While some of the anti-IgE molecules 
      are inactivated during adsorption, the remaining molecules, once adsorbed, appear 
      to retain activity for extended periods, and the coated tubes were stable for 
      several weeks. Kinetic studies of the association of analyte, of antibody-enzyme 
      conjugate and enzymic catalysis were used for the optimisation of the assay and 
      allowed us to reduce assay time to 6 h. The association of analyte to the coated 
      tube and the association of anti-IgE-peroxidase conjugate to bound IgE, in the 
      present design, had T1/2 of approximately 0.5 h. Equilibrium is not obtained. The 
      dissociation of analyte, surprisingly, was nearly undetectable. At least one of 
      the two antibodies employed in the sandwich assay, preferably the 
      anti-IgE-peroxidase conjugate, should be free of non-specific antibodies in order 
      to eliminate non-specific reactions. The influence of serum matrix was eliminated 
      by diluting serum samples and standards 1:21 in buffer. For the assay of very low 
      concentrations (i.e. less than 10 kIU/l) standards produced in IgE-free serum are 
      required to compensate the influence of matrix. Comparison with a commercial kit 
      (Hoechst-Behringwerke, Frankfurt/Main, FRG) showed good agreement. All reagents 
      are commercially available and the assay is well suited for routine use.
FAU - Solling, H
AU  - Solling H
FAU - Dinesen, B
AU  - Dinesen B
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PL  - Netherlands
TA  - Clin Chim Acta
JT  - Clinica chimica acta; international journal of clinical chemistry
JID - 1302422
RN  - 0 (Polystyrenes)
RN  - 0 (Reagent Kits, Diagnostic)
RN  - 37341-29-0 (Immunoglobulin E)
RN  - EC 1.11.1.- (Peroxidases)
SB  - IM
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Humans
MH  - Immunoglobulin E/*analysis
MH  - Peroxidases
MH  - Polystyrenes
MH  - Reagent Kits, Diagnostic
EDAT- 1983/05/09 00:00
MHDA- 1983/05/09 00:01
CRDT- 1983/05/09 00:00
PHST- 1983/05/09 00:00 [pubmed]
PHST- 1983/05/09 00:01 [medline]
PHST- 1983/05/09 00:00 [entrez]
AID - 0009-8981(83)90260-7 [pii]
AID - 10.1016/0009-8981(83)90260-7 [doi]
PST - ppublish
SO  - Clin Chim Acta. 1983 May 9;130(1):71-83. doi: 10.1016/0009-8981(83)90260-7.