PMID- 9261963
OWN - NLM
STAT- MEDLINE
DCOM- 19970926
LR  - 20190831
IS  - 0165-2427 (Print)
IS  - 0165-2427 (Linking)
VI  - 57
IP  - 3-4
DP  - 1997 Jul
TI  - Quantification of total sheep IgE concentration using anti-ovine IgE monoclonal 
      antibodies in an enzyme immunoassay.
PG  - 253-65
AB  - Monoclonal antibodies (mAbs) which recognize separate epitopes on ovine 
      immunoglobulin E (IgE) have been used to develop a non-competitive antibody 
      sandwich enzyme immunoassay (EIA) for quantitating ovine IgE. Purified anti-IgE 
      mAb (YD3) coated onto polystyrene microtitre plates was used to capture IgE in 
      serum samples. Biotinylated anti-IgE mAb (XB6) followed by streptavidin 
      conjugated with horseradish peroxidase were used to detect captured IgE. 
      Tetramethylbenzidine and H2O2 were used as enzyme substrate. A reference serum 
      was prepared by pooling sheep sera containing elevated IgE levels. This reference 
      serum was assigned a value of 100 units ml-1 and used to prepare standard curves 
      for the EIA. The linear region of log-log transformed standard curve data covered 
      a range of 0.05-0.8 units ml-1. The equation of a linear regression line fitted 
      to this curve was used to determine sample concentrations. Using purified IgE, 1 
      unit of reference serum was equivalent to 0.86 micrograms ml-1 IgE. Maximum 
      intra- and inter-assay coefficients of variation for the EIA were 4.6% and 9.7%, 
      respectively. Subjecting serum samples to 15 freeze/thaw cycles, storage at room 
      temperature for 16 days or incubation at 37 degrees C for 8 h resulted in minimal 
      loss of IgE detection. Incubation of serum at 56 degrees C resulted in rapid 
      reduction in detection of IgE by the EIA. The assay was used to determine IgE 
      levels in adult sheep monospecifically infected with weekly doses of the nematode 
      Trichostrongylus axei. Serum IgE levels increased from 9 to 16 days following 
      first infection and reached maximum levels by days 35-58. Serum IgE responses 
      closely followed IgE positive cell responses in the abomasal mucosa.
FAU - Shaw, R J
AU  - Shaw RJ
AD  - AgResearch, Wallaceville Animal Research Centre, Upper Hutt, New Zealand. 
      shawr@agresearch.cri.nz
FAU - McNeill, M M
AU  - McNeill MM
FAU - Gatehouse, T K
AU  - Gatehouse TK
FAU - Douch, P G
AU  - Douch PG
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - Vet Immunol Immunopathol
JT  - Veterinary immunology and immunopathology
JID - 8002006
RN  - 0 (Antibodies, Anti-Idiotypic)
RN  - 0 (Antibodies, Helminth)
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (anti-IgE antibodies)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - Animals
MH  - Antibodies, Anti-Idiotypic/*pharmacology
MH  - Antibodies, Helminth/pharmacology
MH  - Antibodies, Monoclonal/*pharmacology
MH  - Antibody Specificity
MH  - Drug Stability
MH  - Immunoenzyme Techniques/standards/*veterinary
MH  - Immunoglobulin E/*blood
MH  - Reference Standards
MH  - Reproducibility of Results
MH  - Sheep/*immunology
MH  - Trichostrongylus/immunology
EDAT- 1997/07/01 00:00
MHDA- 1997/07/01 00:01
CRDT- 1997/07/01 00:00
PHST- 1997/07/01 00:00 [pubmed]
PHST- 1997/07/01 00:01 [medline]
PHST- 1997/07/01 00:00 [entrez]
AID - S0165-2427(97)00010-X [pii]
AID - 10.1016/s0165-2427(97)00010-x [doi]
PST - ppublish
SO  - Vet Immunol Immunopathol. 1997 Jul;57(3-4):253-65. doi: 
      10.1016/s0165-2427(97)00010-x.