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However, specialized equipment such as FPLC is typically needed but not often available in many laboratories. Here, we show a novel method utilizing polyphosphate (polyP) for purifying proteins with histidine repeats via non\u2010covalent interactions. Our study demonstrates that immobilized polyP efficiently binds to histidine\u2010tagged proteins across a pH range of 5.5\u20137.5, maintaining binding efficacy even in the presence of reducing agent DTT and chelating agent EDTA. We carried out experiments of purifying various proteins from cell lysates and fractions post\u2010Ni\u2010NTA. Our results demonstrate that polyP resin is capable of further purification post\u2010Ni\u2010NTA without the need for specialized equipment and without compromising protein activity. This cost\u2010effective and convenient method offers a viable approach as a complementary approach to Ni\u2010NTA.<\/jats:p>","DOI":"10.1002\/pro.5021","type":"journal-article","created":{"date-parts":[[2024,5,15]],"date-time":"2024-05-15T11:09:30Z","timestamp":1715771370000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Protein purification via consecutive histidine\u2013polyphosphate interaction"],"prefix":"10.1002","volume":"33","author":[{"given":"Zihao","family":"Zhou","sequence":"first","affiliation":[{"name":"School of Pharmaceutical Sciences Central South University  Changsha Hunan China"},{"name":"Department of Biomedical and Molecular Sciences Queen's University  Kingston Ontario Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jin","family":"Jin","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Molecular Sciences Queen's University  Kingston Ontario Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xu","family":"Deng","sequence":"additional","affiliation":[{"name":"School of Pharmaceutical Sciences Central South University  Changsha Hunan China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9322-9394","authenticated-orcid":false,"given":"Zongchao","family":"Jia","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Molecular Sciences Queen's University  Kingston Ontario Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,5,15]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)54198-3"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0291-151"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2015.02.010"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2018.02.104"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-09262-6"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi1014437"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1182\/blood-2011-07-368811"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1160\/TH14-06-0515"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1385\/0-89603-258-2:371"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhazmat.2021.125928"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.f.202"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1042\/BJ20061542"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi034804d"},{"key":"e_1_2_8_15_1","volume-title":"Bioconjugate techniques","author":"Hermanson GT","year":"2013"},{"issue":"9","key":"e_1_2_8_16_1","first-page":"1180","article-title":"Inorganic polyphosphate and specific induction of apoptosis in human plasma cells","volume":"91","author":"Hernandez\u2010Ruiz L","year":"2006","journal-title":"Haematologica"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9673(00)93969-4"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1188-1321"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1523\/JNEUROSCI.0623-07.2007"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.biochem.68.1.89"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12860-019-0221-4"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.266320"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1182\/bloodadvances.2021004567"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.actbio.2009.12.055"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1538-7836.2009.03723.x"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2023.113082"},{"key":"e_1_2_8_27_1","first-page":"139","volume-title":"Lipase and phospholipase activity methods for marine organisms. 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