PMID- 18635267
OWN - NLM
STAT- MEDLINE
DCOM- 20081223
LR  - 20080922
IS  - 0165-2427 (Print)
IS  - 0165-2427 (Linking)
VI  - 125
IP  - 3-4
DP  - 2008 Oct 15
TI  - Porcine IgE in the context of experimental food allergy: purification and 
      isotype-specific antibodies.
PG  - 303-14
LID - 10.1016/j.vetimm.2008.05.028 [doi]
AB  - Measurement of allergen-specific immunoglobulin E (IgE) is a common practice in 
      the investigation of allergy. It has not been possible to measure porcine IgE due 
      to unavailability of anti-porcine IgE. This study was undertaken to purify and 
      characterize porcine IgE from sera of allergic pigs, identify heterologous 
      anti-IgE reactive with pig IgE and to use purified heavy (H) chain of porcine IgE 
      to generate rabbit anti-IgE. A four-step process for the purification of porcine 
      IgE is reported using ammonium sulphate precipitation, Protein G affinity 
      chromatography, DEAE cellulose anion-exchange chromatography and sodium dodecyl 
      sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) to obtain IgE H chain. The 
      resultant IgE was evaluated for purity using SDS-PAGE and immunoreactivity was 
      detected by Prausnitz-Kustner (PK) tests and passive cutaneous anaphylaxis with 
      the allergen, crude peanut extract, used to induce experimental allergy. 
      Cross-reactivity with anti-mouse and anti-human IgE antibodies were confirmed in 
      western blot and enzyme-linked immunosorbent assays (ELISA). The H chain of IgE 
      was excised from SDS-PAGE gels and used to develop rabbit anti-porcine IgE 
      antisera. Antiserum obtained from rabbits immunized with porcine IgE, as well as 
      heterologous murine and human-specific anti-IgE, induced reverse cutaneous 
      anaphylaxis in pig skin and detected allergen-specific IgE in ELISA but did not 
      react with IgG H chain in western blots. These results confirm allergy-associated 
      bioactivity of porcine IgE and describe both homologous and heterologous anti-pig 
      IgE suitable for use in allergen-specific and other assays. This will enhance 
      utility of pig allergy models and provide an additional measure of type-2 immune 
      response in pigs.
FAU - Rupa, Prithy
AU  - Rupa P
AD  - Department of Pathobiology, Ontario Veterinary College, University of Guelph, 
      Guelph, Ontario, Canada.
FAU - Hamilton, Korinne
AU  - Hamilton K
FAU - Cirinna, Melissa
AU  - Cirinna M
FAU - Wilkie, Bruce N
AU  - Wilkie BN
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20080608
PL  - Netherlands
TA  - Vet Immunol Immunopathol
JT  - Veterinary immunology and immunopathology
JID - 8002006
RN  - 0 (Immunoglobulin Heavy Chains)
RN  - 0 (Immunoglobulin Isotypes)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - Animals
MH  - Animals, Suckling
MH  - Antibody Specificity
MH  - Blotting, Western
MH  - Chromatography, Affinity
MH  - Chromatography, Ion Exchange
MH  - Disease Models, Animal
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Immunoglobulin E/blood/*immunology/isolation & purification
MH  - Immunoglobulin Heavy Chains/immunology
MH  - Immunoglobulin Isotypes/*immunology
MH  - Passive Cutaneous Anaphylaxis/immunology
MH  - Peanut Hypersensitivity/*immunology
MH  - Random Allocation
MH  - Specific Pathogen-Free Organisms
MH  - Swine
EDAT- 2008/07/19 09:00
MHDA- 2008/12/24 09:00
CRDT- 2008/07/19 09:00
PHST- 2008/04/01 00:00 [received]
PHST- 2008/05/21 00:00 [revised]
PHST- 2008/05/30 00:00 [accepted]
PHST- 2008/07/19 09:00 [pubmed]
PHST- 2008/12/24 09:00 [medline]
PHST- 2008/07/19 09:00 [entrez]
AID - S0165-2427(08)00255-9 [pii]
AID - 10.1016/j.vetimm.2008.05.028 [doi]
PST - ppublish
SO  - Vet Immunol Immunopathol. 2008 Oct 15;125(3-4):303-14. doi: 
      10.1016/j.vetimm.2008.05.028. Epub 2008 Jun 8.