PMID- 9656428
OWN - NLM
STAT- MEDLINE
DCOM- 19980923
LR  - 20190831
IS  - 0165-2427 (Print)
IS  - 0165-2427 (Linking)
VI  - 64
IP  - 1
DP  - 1998 Jun 30
TI  - Characterization of two dog IgE-specific antibodies elicited by different 
      recombinant fragments of the epsilon chain in hens.
PG  - 15-32
AB  - Two recombinant [His]6-tagged fragments of the canine immunoglobulin E (IgE) 
      heavy chain (second domain: IgEf2 and third and fourth domains: IgEf3/4) were 
      cloned, expressed in Escherichia coli (E. coli) as [His]6-tagged proteins, and 
      affinity-purified over nickel-nitrilotriacetic acid columns. The recombinant 
      proteins were used to immunize hens. The raised and affinity-purified chicken 
      antibodies (Ab) isolated from egg yolk exhibited specific binding to the 
      respective recombinant canine IgE fragment (IgEf) on immunoblots and displayed 
      high titers against the IgEf in ELISA. Immunoblotting of canine serum separated 
      by PAGE under native conditions with the IgEf2- and IgEf3/4-specific Ab resulted 
      in staining of a protein of approximately 180 kilodaltons (kD). The 
      IgEf3/4-specific Ab further recognized an 80 kD protein in IgEf3/4-specific Ab 
      affinity-enriched dog serum separated under denaturing conditions. In an ELISA 
      for the detection of antigen-specific IgE in dog serum, reduced binding of the 
      IgEf-specific Ab was observed after heat treatment of the dog serum. The 
      reactivity of both of the raised chicken Ab was only present in postimmune 
      reagents and could only be inhibited by preincubation with the IgEf used for 
      immunization and not with dog immunoglobulin G, E. coli extract, or with a 
      nonrelevant recombinant [His]6-tagged protein. In immunohistochemistry, the 
      IgEf3/4-specific Ab specifically recognized cells in paraffin-embedded tissue 
      sections of lymph nodes. Furthermore, both of the IgEf-specific Ab elicited 
      positive immediate type 1 skin reactions in dogs. Semiquantitative assessment of 
      total serum IgE in dogs was developed using IgEf2-specific Ab as coating reagent 
      and the biotinylated IgEf3/4-specific Ab as developing Ab in ELISA. In 
      conclusion, both IgEf-specific Ab recognize native dog IgE with the advantages 
      that they are directed against different and known constant domains of the IgE 
      molecule, and that they can be used for immunohistochemistry on paraffin-embedded 
      tissue. The two dog IgE-specific Ab could initiate clinical research on the 
      involvement of immediate-type hypersensitivity reactions in dogs.
FAU - Griot-Wenk, M E
AU  - Griot-Wenk ME
AD  - Institute of Animal Breeding, Division of Immunogenetics, Bern, Switzerland. 
      monika.griot@itpa.unibe.ch
FAU - Marti, E
AU  - Marti E
FAU - Racine, B
AU  - Racine B
FAU - Crameri, R
AU  - Crameri R
FAU - Zurbriggen, A
AU  - Zurbriggen A
FAU - de Weck, A L
AU  - de Weck AL
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 (Antibodies, Anti-Idiotypic)
RN  - 0 (DNA Primers)
RN  - 0 (Immunoglobulin Fragments)
RN  - 0 (Immunoglobulin epsilon-Chains)
RN  - 0 (Recombinant Proteins)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - Animals
MH  - Antibodies, Anti-Idiotypic/*biosynthesis/immunology/isolation & purification
MH  - Antibody Specificity
MH  - Base Sequence
MH  - Chickens/*immunology
MH  - Cloning, Molecular
MH  - DNA Primers/genetics
MH  - Dogs/*immunology
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Escherichia coli/genetics
MH  - Female
MH  - Immunization
MH  - Immunoglobulin E/analysis/*immunology
MH  - Immunoglobulin Fragments/chemistry/genetics/immunology
MH  - Immunoglobulin epsilon-Chains/chemistry/genetics/*immunology
MH  - Polymerase Chain Reaction
MH  - Recombinant Proteins/chemistry/genetics/immunology
MH  - Species Specificity
EDAT- 1998/07/10 00:00
MHDA- 1998/07/10 00:01
CRDT- 1998/07/10 00:00
PHST- 1998/07/10 00:00 [pubmed]
PHST- 1998/07/10 00:01 [medline]
PHST- 1998/07/10 00:00 [entrez]
AID - S0165-2427(98)00118-4 [pii]
AID - 10.1016/s0165-2427(98)00118-4 [doi]
PST - ppublish
SO  - Vet Immunol Immunopathol. 1998 Jun 30;64(1):15-32. doi: 
      10.1016/s0165-2427(98)00118-4.