Epitope mapping of SHP-1 monoclonal antibodies using peptide phage display

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摘要：

We have characterized the of four monoclonal antibodies for , an domain containing protein tyrosine , using two phage displayed random libraries. Three of the antibodies are directed against the domain of the molecule and the fourth is toward the -terminal part of the second domain. The first two antibodies recognize the sequence NANY, amino acid 305 to amino acid 308, numbered in the non haematopoietic form of sequence. The third antibody binds the sequence P Y W P (amino acids 365 to 368) located toward the middle of the domain of the enzyme. The fourth antibody is directed against the first two amino acids, W Y (amino acids 112 and 113), of the second domain. The specificities of these antibodies are demonstrated by ELISA and western blot using different protein constructs expressed in . All the antibodies can detect wild type , expressed in 293 cells, by western blot analysis, both under denaturing conditions as well as following renaturation. The data presented here show that the antibodies characterized in this study are raised against linear and suggest that these are accessible from the outside in the native molecule.

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关键词：

Humans Bacteriophages Intracellular Signaling Peptides and Proteins Antibodies, Monoclonal Blotting, Western Enzyme-Linked Immunosorbent Assay Epitope Mapping src Homology Domains Antibody Specificity Amino Acid Sequence