Use of Seprion to identify abnormal aggregated proteins in Alzheimer’s Disease brain

Samples.



Aggregated proteins were first extracted by Seprion followed by ELISA for the proteins of interest.

 

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Use of Seprion to monitor the development with age of ß-amyloid aggregates in an Alzheimer’s mouse model.


Brains of mice at different ages were homogenised, extracted with Seprion and then tested in ELISA.


The increase of ß-amyloid aggregates can be seen with age.

 

Use of Seprion to extract the abnormal prion protein from brain and spleen  followed by an ELISA specific for the prion protein.




Seprion extracted the prion aggregates from multiple species and organs.


Signal

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References

1. S. Wilson et al. 2004. A protein-binding ligand system for prion disease detection. IVD Technology September 2004.

2. K. Sathasivam et al. Identical oligomeric and fibrillar structures captured from the brains of R6/2 and knock-in mouse models of Huntington's disease. 2010; Human Mol. Gen. 19(1):65-78.

3. L. A. Terry, et al. Detection of PrPsc in Blood from Sheep Infected with the Scrapie and Bovine Spongiform Encephalopathy Agents. 2009. J. Virol. 83(23): 12552-12558.


Use of Seprion to detect aggregated IgG and the influence of size-filtration.


Aggregated IgG was extracted using Seprion and then detected by ELISA.


Aggregates were detected but removed by filtration (300 KDa molecular weight cut-off).


Fractions:

1, Non-aggregated, not filtered

2. Non-aggregated but filtered

3. Aggregated, not filtered

4. Aggregated, filtered

The Seprion ligand is a unique versatile tool that can be used to separate abnormal aggregated proteins from the normal unaggregated protein (ref. 1) Seprion has been used in the study of a wide range of protein aggregation diseases and in the monitoring of aggregation in the production of therapeutic proteins. Proteins studied include: aggregated therapeutic antibodies; huntingtin in Huntington’s Disease in animal models (ref. 2); β-amyloid and tau in Alzheimer’s Disease in both human disease and in animal models; α-synuclein in Parkinson’s Disease; abnormal prion protein in both sporadic and new variant Creutzfeldt-Jakob Disease (CJD); Bovine Spongiform Encephalopathy (BSE) in cows; Chronic Wasting Disease (CWD) in deer; scrapie, including atypical scrapie in sheep (ref. 3). Kits based on Seprion have USDA and EU approval for animal testing (the Idexx Herdchek Test). The Seprion ligand is used by the Health Protection Agency, UK in their tonsil screening programme and worldwide for food safety in the screening of millions of cattle for BSE.

See examples of the range of uses of Seprion below:

Aggregation  2011/1