Dataset Open Access

Large-scale stringent purification of Q23 HTT and HTT-HAP40 using heparin affinity chromatography 2019/05/22

Harding, Rachel; Alvarez, Claudia; McAuley, Jacob; Hutchinson, Ashley; Seitova, Alma; Arrowsmith, Cheryl; Edwards, Aled


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    <subfield code="a">Dr. Harding is the recipient of the Huntington's Disease Society of America Berman Topper Career Development Fellowship which funds and supports this research, in addition to generous funding from the CHDI Foundation and the Huntington Society of Canada. 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.</subfield>
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    <subfield code="a">&lt;p&gt;&lt;strong&gt;Project:&amp;nbsp;&lt;/strong&gt;Biophysical investigation of purified HTT protein samples&lt;/p&gt;

&lt;p&gt;&lt;strong&gt;Experiment:&amp;nbsp;&lt;/strong&gt;Large-scale stringent purification of Q23 HTT and HTT-HAP40 using heparin affinity chromatography&lt;/p&gt;

&lt;p&gt;&lt;strong&gt;Date completed:&amp;shy;&amp;nbsp;&lt;/strong&gt;2019/05/22&lt;/p&gt;

&lt;p&gt;&lt;strong&gt;Rationale:&amp;nbsp;&lt;/strong&gt;Previous attempts to generate a much purer and homogenous HTT-HAP40 sample showed that the complex can bind heparin resin. In small-scale experiment, I also incorporated helpful suggestions from scientists at the CHDI Palm Springs meeting full-length HTT research breakout group i.e. ATP wash to remove HSP proteins. The experiment included 3 affinity chromatography steps with FLAG, heparin and NiNTA resin and finally a gel filtration step and yielded a highly pure HTT-HAP40 sample:&amp;nbsp;https://zenodo.org/record/3234174.I now want to try this purification with apo HTT.&amp;nbsp;&lt;/p&gt;</subfield>
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