| Products/Services Used | Details | Operation |
|---|---|---|
| Peptides were synthesized by standard chemical peptide synthesis and purity was analyzed by HPLC (GenScript, NJ).The antibody fragments of VH, VL, CH and CL were amplified according to the standard operating procedure for RACE at GenScript (Genscript, Piscataway, NJ). 1C9 antibody was humanized by Genscript using proprietary technology.High tau protein-binding phages were selected after three rounds of panning using Genscript’s proprietary FASEBA screening methodology. Filtered supernatant was loaded onto Protein A CIP column 65 ml (GenScript, Cat.No.L00433) at 8.0 ml/min. After washing with PBS, elution with 50 mM Citric acid, pH 3.0 and neutralization with 1 M Tris–HCl, pH 9.0, the eluted fractions were desalted by HiPrep 26/10 Desalting column with PBS.96-well plates (Immulux HB; Dynex Technologies, Inc., VA) were coated with 1 μg/well (in 100 μl; Carbonate-Bicarbonate buffer, pH 9.6, o/n at 4 °C) tau2– 18 peptide (GenScript, NJ). |
The experience from clinical trials indicates that anti-Aβ immunotherapy could be effective in early/pre-clinical stages of AD， whereas at the late stages promoting the clearing of Aβ alone may be insufficient to halt the disease progression. At the same time， pathological tau correlates much better with the degree of dementia than Aβ deposition. Therefore， targeting pathological tau may provide a more promising approach for the treatment of advanced stages of AD. Recent data demonstrates that the N-terminal region of tau spanning aa 2-18 termed "phosphatase activation domain" that is normally hidden in the native protein in 'paperclip'-like conformation， becomes exposed in pathological tau and plays... More
