PMID- 9477476
OWN - NLM
STAT- MEDLINE
DCOM- 19980326
LR  - 20190831
IS  - 0165-2427 (Print)
IS  - 0165-2427 (Linking)
VI  - 59
IP  - 3-4
DP  - 1997 Nov
TI  - Chicken antibodies to a recombinant fragment of the equine immunoglobulin epsilon 
      heavy-chain recognising native horse IgE.
PG  - 253-70
AB  - An equine immunoglobulin E (IgE) heavy-chain cDNA fragment (CH3-CH4, nucleotides 
      1132 to 1592) was cloned, expressed in Escherichia coli as a fusion protein with 
      a [His]6-tag and purified over a Ni-NTA column. The recombinant protein was used 
      to immunise hens. Testing of the raised egg yolk immunoglobulin G (IgG) in 
      Western-blot and ELISA revealed high titres against the recombinant equine IgE 
      fragment (reqIgEf). The reqIgEf-specific IgG was successfully affinity-purified 
      on an unconventional affinity matrix: the [His]6-tagged recombinant IgE fragment 
      was bound to Ni-NTA agarose and used to adsorb specific immunoglobulins. In 
      Western-blot of ammonium sulphate precipitated horse serum and bronchoalveolar 
      lavage fluid, separated by SDS-PAGE under denaturing-reducing conditions, the 
      raised antibodies reacted with a protein of approximately 80 kDa. A reaction of 
      the reqIgEf-specific IgG was seen with a 190-200 kDa band when the same horse 
      serum or bronchoalveolar fluid (BALF) was separated under non-reducing 
      conditions. These reactions could be inhibited by preincubation of the immune IgG 
      with reqIgEf, while preincubation with horse IgG did not inhibit the reaction. 
      Antibody-affinity chromatography of horse serum with the reqIgEf-specific chicken 
      IgG resulted in an enrichment of the 80 kDa protein in denaturing Western-blot. 
      Determination of the amino acid composition of this protein and comparison with 
      the equine IgE heavy- chain sequence strongly indicates that the 80 kDa band 
      corresponds to the heavy chain of the horse IgE. The reqIgEf-specific chicken IgG 
      was further characterised in an ELISA for the detection of allergen-specific 
      horse IgE. It was demonstrated that heating IgE positive horse sera at 54 degrees 
      C for 10 min drastically diminished the recognition by the reqIgEf-specific 
      chicken IgG. The reaction is inhibitable by preincubation with reqIgEf in a 
      concentration dependent manner. In addition, preincubation with horse IgG, a 
      nonrelevant [His]6-tagged protein or 2% equine colostrum had no influence on the 
      reqIgEf-specific chicken IgG binding characteristic. This antibody recognising 
      horse IgE will be useful for further studies on the pathogenesis of equine 
      allergic diseases.
FAU - Marti, E
AU  - Marti E
AD  - Klinik fur Nutztiere und Pferde, University of Berne, Switzerland. 
      marti@itz.unibe.ch
FAU - Peveri, P
AU  - Peveri P
FAU - Griot-Wenk, M
AU  - Griot-Wenk M
FAU - Muntwyler, J
AU  - Muntwyler J
FAU - Crameri, R
AU  - Crameri R
FAU - Schaller, J
AU  - Schaller J
FAU - Gerber, H
AU  - Gerber H
FAU - Lazary, S
AU  - Lazary S
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 (Amino Acids)
RN  - 0 (Antibodies, Anti-Idiotypic)
RN  - 0 (Immunoglobulin epsilon-Chains)
RN  - 0 (Indicators and Reagents)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - Amino Acids/analysis
MH  - Anaphylaxis/immunology/veterinary
MH  - Animals
MH  - Antibodies, Anti-Idiotypic/*immunology
MH  - Chickens
MH  - Chromatography, Affinity
MH  - Cloning, Molecular
MH  - Colostrum/immunology
MH  - Escherichia coli
MH  - Goats
MH  - Horses
MH  - Hot Temperature
MH  - Immunoblotting
MH  - Immunoglobulin E/*immunology
MH  - Immunoglobulin epsilon-Chains/genetics/*immunology/isolation & purification
MH  - Indicators and Reagents
MH  - Rabbits
MH  - Recombinant Fusion Proteins/genetics/immunology/isolation & purification
EDAT- 1998/02/27 00:00
MHDA- 1998/02/27 00:01
CRDT- 1998/02/27 00:00
PHST- 1998/02/27 00:00 [pubmed]
PHST- 1998/02/27 00:01 [medline]
PHST- 1998/02/27 00:00 [entrez]
AID - S0165-2427(97)00096-2 [pii]
AID - 10.1016/s0165-2427(97)00096-2 [doi]
PST - ppublish
SO  - Vet Immunol Immunopathol. 1997 Nov;59(3-4):253-70. doi: 
      10.1016/s0165-2427(97)00096-2.