ABSTRACT
Amyloid beta-protein (Abeta) has been causally implicated in the neurodegenerative processes that accompany Alzheimer's disease. Soluble oligomers of the Abeta(1-42) fragment are thought to be significantly more neurotoxic than higher molecular weight aggregates. We report the isolation and characterization of a mouse monoclonal antibody (MAb) directed against soluble Abeta(1-42) oligomers. Synthetic Abeta(1-42) oligomers were assembled in vitro; these were used to immunize mice, and hybridomas were isolated following myeloma fusion of splenocytes from immunized animals. Screening for reactivity against Abeta(1-42) resulted in the identification of MAb A8 with high affinity for soluble oligomers. The isotype of A8 was found to be IgG(2b). Experiments using sub-peptides of Abeta(1-42) revealed that the epitope identified by A8 lies within the 1-6 region of Abeta. The antibody displays high affinity for soluble Abeta(1-42) oligomers in the molecular weight range of 16.5-25 kDa, and detected target antigen in brain sections from senescence-accelerated SAMP 8 mice. The sensitivity and optimal titers for the detection of soluble Abeta(1-42) oligomers were determined to be 0.625 microg/mL in indirect ELISA, and 1:10(6), 1:4000, and 1:150 for ELISA, Western blotting, and immunohistochemistry, respectively. The A8 antibody specific for soluble Abeta(1-42) oligomers will provide a valuable tool for Alzheimer's disease research.
Subject(s)
Amyloid beta-Peptides/immunology , Antibodies, Monoclonal/biosynthesis , Amino Acid Sequence , Animals , Antibodies, Monoclonal/chemistry , Antibodies, Monoclonal/immunology , Antibody Affinity , Antibody Specificity , Blotting, Western , Electrophoresis, Polyacrylamide Gel , Enzyme-Linked Immunosorbent Assay , Female , Immunohistochemistry , Mice , Mice, Inbred BALB C , Molecular Sequence DataABSTRACT
@#Objective To explore the management of deep vein thrombosis (DVT) of lower limbs in patients with severe craniocerebral injury.Methods The clinical data of 9 patients of severe craniocerebral injury with DVT were analyzed respectively.Results All 9 patients were given medicine therapy including thrombolytic, anti coagulating, anti platelet aggregation and antibiotics. 3 cases were cured, 1 case was improved, 4 cases died and 1 case discharged by himself. Conclusion There are risk factors for DVT in patients with severe craniocerebral injury. Early prophylaxis is important. Early diagnosis and treatment are benefited.
