Monoclonal antibodies to human myelin basic protein.
SJL/J and (SJL X PL) F1 hybrid mice were immunized with intact human myelin basic protein (MBP) or the three major peptic fragments of MBP, residues 1-38, 39-89, and 90-170. Immune spleen cells were fused with mouse myeloma P3 X 63Ag8 (NS1) cells in the presence of polyethylene glycol. Hybridoma supernatant culture fluids were screened for antibody to MBP by a solid-phase radioimmunoassay (RIA). The specificity of the monoclonal antibody (mAb) was characterized by RIA using the three major MBP peptic fragments and subfragments as well as MBP and MBP fragments of different species with known amino acid sequence differences. Six MBP mAbs were generated, one of them IgM isotype and the remainder IgG isotypes. One mAb each reacted against regions of residues 22-38, 39-69, 70-89, 90-116, and two reacted against residues 118-157. Immunoblots also showed that the five IgG mAbs were reactive against MBP and the peptic fragment of MBP containing the epitope. Immunohistochemical studies showed the IgG mAbs specifically stained myelinated fiber tracts in human brain tissue.
Chou
C H
CH
Cox
A A
AA
Fritz
R B
RB
Wood
J G
JG
Kibler
R F
RF
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D000939:Epitopes / Q000276:immunology; D006168:Guinea Pigs; D006801:Humans; D051379:Mice / Q000276:immunology; D004676:Myelin Basic Protein / Q000235:genetics / Q000276:immunology*; D011863:Radioimmunoassay; D051381:Rats
D000911:Antibodies, Monoclonal; D000939:Epitopes; D004676:Myelin Basic Protein