Monoclonal and polyclonal antibody responses to the myelin basic protein epitope present in human urine.
The myelin basic protein (MBP)-like material (MBPLM) in human urine is expressed in a cryptic epitope(s) present in MBP peptide 80-89 and absent or inaccessible in intact MBP. These features of urinary MBPLM have made it difficult to produce reagents for its further characterization. Using an immunogen of keyhole limpet hemocyanin conjugated to MBP peptide cys 74-89, polyclonal antiserum to urinary MBPLM was prepared. With the same immunogen and screening with urine, the product from one of over 1600 wells from the original fusion, produced monoclonal antibody (mAb) which could detect urinary MBPLM. By radioimmunoassay two rabbit polyclonal reagents recognized a cryptic epitope in MBP peptide 84-89 while the two mAbs recognized another cryptic epitope in MBP peptide 80-85. Both could be used for quantitation of MBPLM in urine. These and previous results indicate the presence of at least three epitopes, one noncryptic and two cryptic, in the decapeptide of MBP 80-89. Of the two cryptic epitopes, the one near the carboxyl-terminal is dominant to that in the amino-terminal portion. The detection of urinary MBPLM with reagents with two different specificities suggests the presence of two or more small MBP peptides in urine.
Whitaker
J N
JN
Department of Neurology, University of Alabama at Birmingham 35294-0007.
McKeehan
A
A
Freeman
D W
DW
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000939:Epitopes; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D004676:Myelin Basic Protein / Q000276:immunology* / Q000652:urine; D011817:Rabbits
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000939:Epitopes; D004676:Myelin Basic Protein