Polypeptide antigen and antibody related to Tau protein

An antibody and protein technology, applied in the direction of anti-animal/human immunoglobulin, peptide source, animal/human peptide, etc.

Inactive Publication Date: 2006-01-18
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, whether protein isoform 6d exists in the form of protein cannot be directly concluded from the above published literature

Method used

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  • Polypeptide antigen and antibody related to Tau protein
  • Polypeptide antigen and antibody related to Tau protein
  • Polypeptide antigen and antibody related to Tau protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Preparation of polypeptide antigen

[0025] The polypeptide antigen provided by the present invention has the amino acid sequence described in SEQ ID No.1, namely FWSKGDETQGG. Can be synthesized by commercial companies. In this application, the polypeptide antigen was synthesized, purified, and freeze-dried by Zymed Company, and the results are shown in figure 2 , 3 .

Embodiment 2

[0027] Preparation of specific polyclonal antibody (6d) of Tau protein isotype 6d:

[0028] The antigen obtained above was conjugated to KLH [5] After that, immunize animals (rabbits) [6，7] Preparation of antiserum, purification of antiserum to obtain 6d antibody [8，9] , the purified 6d antibody was separated by 10% SDS-PAGE and detected by Coomassi staining ( Figure 4 ) [10] .

[0029] (1) Immunized animals (rabbits) [6，7] Prepare antiserum:

[0030] Add an equal amount of complete Freund's adjuvant to 1 ml of polypeptide antigen solution (1 mg / ml, dissolved in PBS), and emulsify with a double-tube micro-emulsification needle (Thomas Scientific 345OD46). The hair on the back of the rabbit was shaved and disinfected with 70% alcohol. Intradermal injection was performed with a 3ml syringe (24-G) needle at an angle of 30° to the skin, and the injection was divided into 24 points, each injection point was injected with 25 μl, and a total of 3 rabbits were immunized. Six ...

Embodiment 3

[0041] The 6d antibody is used to detect and identify whether Tau protein isoform 6d exists in the form of protein: use 6d as the primary antibody, and detect the expression level and spatiotemporal distribution of endogenous Tau protein isoform 6d in different parts of the brain tissue by Western Blotting; The Tau protein isoform 6d gene was cloned and introduced into COS cells for in vitro expression, and the exogenous Tau protein isoform 6d was detected and identified by Western Blotting. The results showed that A. The new Tau protein isoform 6d did exist in the form of protein ( Figure 5 ,6);

[0042] B. The prepared 6d antibody is a highly specific antibody ( Figure 5 , 6) [11] .

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Abstract

The present invention relates to Tau protein relative polypeptide antigen and antibody. The specific polypeptide antigen of the present invention has the amino acid sequence of SEQ ID No. 1, and corresponding polyclonal or monoclonal antibody may be obtained in available method. The antibody of the present invention may be prepared into kit for detecting and diagnosing senile dementia.

Description

technical field [0001] The invention relates to polypeptide antigens and antibodies related to Tau protein. technical background [0002] The microtubule system is a component of the neurocytoskeleton that is primarily involved in a variety of cellular functions. Microtubules are composed of tubulin and microtubule-associated proteins, and Tau protein is the microtubule-associated protein. The function of normal Tau protein is to combine with tubulin to promote its polymerization to form microtubules; bind with the formed microtubules to maintain the stability of microtubules and reduce the dissociation of tubulin molecules. The Tau protein gene is located on the long arm of chromosome 17. Normal people can express at least 6 isotypes due to the different editing methods of Tau protein RNA. Tau protein is a phosphate-containing protein, and the tau protein molecule in the normal mature brain contains 2 to 3 phosphate groups. In AD patients, the Tau protein is abnormally ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K16/18G01N33/68G01N33/563G01N33/53
Inventor 罗敏华
Owner CENT SOUTH UNIV
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