Antibodies to tau
a technology of antibodies and tau, applied in the field of antibodies to tau, can solve the problems of increasing public health problems, cognitive function loss, and ultimately loss of independence, and the patient and the family take a heavy personal and financial toll
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[0129]The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples that follow represent techniques discovered by the inventors to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention.
Introduction to Examples 1-8
[0130]Aggregation of the microtubule associated protein tau in neurons and glia is associated with over 20 neurodegenerative disorders including Alzheimer disease (AD), progressive supranuclear palsy, and frontotemporal dementia. Recent evidence from human studies suggests that tau pathology does not distribute r...
example 1
Antibodies
[0131]Two series of anti-tau antibodies were created using standard techniques: the HJ8 series (mouse monoclonal antibodies against recombinant human tau), and the HJ9 series (mouse monoclonal antibodies against recombinant mouse tau) (Table 1). Binding epitopes have been mapped for many of the antibodies (Table A).
TABLE 1HJ8 series and HJ9 series against human and mouse tauAntibodyIsotypeApplicationHJ 8.1IgG2b IgG1IP WB IHC(h&m)HJ 8.2IgG2bIP WB IHC(h&m)HJ 8.3IgG2bIP WB IHC(h&m)HJ 8.4IgG1IP WB IHC(h&m)HJ 8.5IgG2bIP WB IHC(h) ELISA forcoating staining 3mon old miceHJ 8.7IgG2bIP WB IHC(h&m) HJ8.7B forELISA detact stainingHJ 8.8IgG2bIP WB IHC(h&m) stainingHJ 9.1IgG2bIP WB IHC ELISAHJ 9.2unknownIP WB IHC ELISA forcoating stainingHJ 9.3IgG2bIP WB IHC ELISA for coatingHJ 9.4IgG2bIP WB IHCHJ 9.5IgG2bIP WB IHCIP = Immunoprecipitation;WB = Western] Blot;ELISA = Enzyme-linked immosorbent Assay,IHC = immunohistochemistry;h = human;m = mouse
[0132]To characterize the binding affinity o...
example 2
th Tau is Present in ISF
[0135]Tau was immunoprecipitated from ISF samples of both wild-type mice and P301 S human tau transgenic mice (P301 S tg mice, details in Methods) using tau antibodies recognizing both mouse and human tau. Two anti-tau monoclonal antibodies that worked well in immunoprecipitation assays were used, as the amount of monomeric tau in ISF is relatively low. Following immunoprecipitation, tau was analyzed by immunoblot. Endogenous murine tau isoforms migrate at 48-62 kDa. In wild-type brain lysate, tau appeared in four separate bands on SDS-PAGE (FIG. 14A). The most abundant species in wild-type mice migrated at 48 kDa. In P301 S tg mice brain, in addition to the four endogenous murine tau bands, overexpressed human 1 N4R tau was observed as an intense band migrating at 55 kDa as well as a 39 kDa band, which may represent a tau degradation product.
[0136]In contrast to total brain lysates, upon immunoprecipitation a single tau band was detected with antibody HJ9.3 ...
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