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US 5,514,776 AGranted
Peptides representing antigenic epitopes of dog IgE present on B cell but not basophil surface
AssigneeTanox Biosystems, Inc.
InventorTse W. Chang
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Abstract
Antigenic epitopes associated with the extracellular segment of the domain which anchors dog immunoglobulin-.epsilon. to the B cell membrane are disclosed. The epitopes are present on dog IgE-bearing B cells but not basophils or the secreted, soluble form of dog IgE. The peptides representing the epitopes associated with the anchor domain of dog IgE can be used to generate antibodies against these regions.
Claims1
A peptide having the sequence of the .epsilon..mb/ec segment of a dog produced by a method comprising:
- preparing the dog genomic library by transfecting it into phage and selecting a phage clone positive for DNA encoding membrane-bound .epsilon. immunoglobulin;
- purifying the phage DNA;
- restriction digesting and analyzing the digested DNA fragments to identify a fragment which includes DNA encoding the membrane-bound domains of .epsilon. immunoglobulin and some of the adjacent constant region;
- subcloning and restriction enzyme mapping said fragment into vectors;
- sequencing said fragment;
- identifying the portion of said fragment coding for membrane .epsilon. immunoglobulin by comparison with a known membrane coding sequence;
- identifying the portion of said fragment coding for the extracellular membrane-bound domain of dog .epsilon. immunoglobulin, based on a plot of the hydrophobicity/hydrophilicity of the amino acid sequence it codes for;
- and
- producing the .epsilon..mb/ec peptide coded for by said portion and isolating said peptide.
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Dependencies are inferred from the claim text.