• This topic has 2 replies, 3 voices, and was last updated 3 years ago by Hüseyin Besir.
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    • March 9, 2022 at 21:04 #4253

      Nikolay Dobrev
      Participant

      Dear all,
      I am looking for someone who can share any experience removing Ferritin contamination when purification from mammalian suspension cells is performed.
      The Ferritin contamination co purifies via gradient for AAV particles. If anyone knows some tricks how to convert the 24-mer Ferritin to a monomeric form ( besides the case with low pH) will be pretty helpful and appreciated.
      Furthermore, if someone is aware of specific affinity resin ( like nanobody) will be great to share.

      Thank you,
      Nikolay

    • March 11, 2022 at 10:03 #4254

      Mario Lebendiker
      Participant

      Hi Nikolay,
      the pI of Ferritin is near 5.5. So, around pH 7.5 8.0 it will possible bind to anion exchange columns more stronger than AAV. You can contact me if you need some advice how to do it.
      I would give this a chance
      Mario

    • April 12, 2022 at 11:37 #4279

      Hüseyin Besir
      Participant

      Hi Nikolay,
      if you haven’t tried Mario’s suggestion, here is a paper describing the separation of ferritin from an AAV preparation using ion exchange. The ferritin seems to bind tightly under these conditions.
      https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4817810/

      Hope that helps.
      Hüseyin

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