Dataset Open Access

Optimisation of HTT-HAP40 purification using heparin affinity chromatography - 2019/01/29

Harding, Rachel; Arrowsmith, Cheryl; Edwards, Aled

Project: Biophysical investigation of purified HTT protein samples

Experiment: Optimisation of HTT-HAP40 purification using heparin affinity chromatography

Date completed:­ 2019/01/29

Rationale: The current protocol for HTT and HTT-HAP40 purification I am using requires a long incubation of clarified cell lysate with FLAG resin. To potentially improve yields and sample quality, it would perhaps be beneficial to have a quick heparin resin purification step prior to FLAG binding which may also remove contaminating nucleic acid material. To test this hypothesis, small-scale purification of Q23 HTT-HAP40 samples in different buffer systems were conducted using heparin and FLAG affinity chromatography which showed the sample bound heparin resin – see https://zenodo.org/record/2553669. Now this need to be scaled up and tested more stringently. 

Dr. Harding is the recipient of the Huntington's Disease Society of America Berman Topper Career Development Fellowship which funds and supports this research. Dr. Harding's research is also supported with generous funding from the Huntington Society of Canada and the CHDI Foundation. The SGC is a registered charity (number 1097737) that receives funds from AbbVie, Bayer Pharma AG, Boehringer Ingelheim, Canada Foundation for Innovation, Eshelman Institute for Innovation, Genome Canada through Ontario Genomics Institute [OGI-055], Innovative Medicines Initiative (EU/EFPIA) [ULTRA-DD grant no. 115766], Janssen, Merck KGaA, Darmstadt, Germany, MSD, Novartis Pharma AG, Ontario Ministry of Research, Innovation and Science (MRIS), Pfizer, São Paulo Research Foundation-FAPESP, Takeda, and Wellcome.
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Publication date:
January 29, 2019
DOI:
10.5281/zenodo.2600366

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Keyword(s):
Huntingtin HTT Huntington's Disease HD HAP40 Heparin
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