Monoclonal antibodies selective for low molecular weight neurofilaments.
Neurofilaments are necessary for the maintenance of axonal caliber and structural organization of nerve cells. The low molecular weight form of neurofilament, the neurofilament light protein, which serves as the core of the filament, was used as immunogen for generation of hybridomas with selective reactivity with this form of the filament. Six hybridomas, out of approximately 100 tested clones, were highly discriminatory. All involved epitopes were localized to the rod region of the antigen, as determined by alpha-chymotrypsin digestion of the purified filament and enzymatic peptide mapping. Synthetic peptides (20 mers) covering the entire rod region did not react with the antibodies. A phage display peptide library was used to identify four consensus sequences for the antibodies. The results indicate that all epitopes were of conformational type. Pair-wise BIAcore data furthermore indicated that the epitopes were independent. The access to such specific reagents is a prerequisite for further elucidation of the biology of the low molecular weight form of neurofilaments proteins.
Norgren
Niklas
N
Department of Immunology, University of Umeå, S-90187 Umeå, Sweden.
Karlsson
Jan-Erik
JE
Rosengren
Lars
L
Stigbrand
Torgny
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D001369:Axons / Q000378:metabolism; D015153:Blotting, Western; D002417:Cattle; D002918:Chymotrypsin / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D000939:Epitopes; D015723:Gene Library; D006825:Hybridomas / Q000378:metabolism; D008251:Macaca; D051379:Mice; D008807:Mice, Inbred BALB C; D009474:Neurons / Q000378:metabolism*; D019151:Peptide Library; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary
D000911:Antibodies, Monoclonal; D000939:Epitopes; D019151:Peptide Library; D010455:Peptides; D002918:Chymotrypsin; C117608:alpha-chymotrypsin