I need large amounts of anti-CD20 and anti-pan-cytokeratin antibodies. At DAKO, they go for >$1,500/mg, and I need >100mg, hence a purchase is out of question.

Instead, I plan to make a homebrew antibody. I have been producing monoclonals from hybridomas and from HEK cells for a decade, and my lab avails of all necessary equipment (Äkta, cell culture, etc.) and expertise.

What I have never done, however, is to start with a published sequence and create (and express in suitable mammalian cells) an artificial antibody H+L gene. Specifically, I was thinking of pulling a sequence from a patent filing, as these tend to be quite precise for legal reasons, making a synthetic gene, and expressing it in HEK.

A nice precedent for that would be for example http://www.ncbi.nlm.nih.gov/pubmed/25451031. Here, the author of the original publication refused to provide (or even sell at any price) the ICSM18 antibody, for reasons that he did not disclose (but which we can all imagine, given that the primary paper was published in Nature and a refutation would be seen as a dishonor). As a result, Colby lifted the sequence from the patent, and made the antibody in their kitchen! (I must say that I love democracy in science!!!)

My question is:

  • might you be able to direct me to good, high-affinity antibodies to CD20 and cytokeratins whose sequences are retrievable? The latter is true only for a minute minority of all available antibodies
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    $\begingroup$ This question needs to be updated with further details. Is there a patent on the antibodies in question? Are the antibodies polyclonal or monoclonal? $\endgroup$ – Michael_A Aug 6 '16 at 10:30
  • $\begingroup$ I am not so sure if you will get functional antibodies this way. $\endgroup$ – Chris Aug 6 '16 at 10:39
  • $\begingroup$ Apologies for the question being vague. I will remedy. $\endgroup$ – aag Aug 6 '16 at 11:17
  • $\begingroup$ Not directly related to the question, but IIRC it's against Nature's publishing policies to deny supplies necessary to replication to other scientists within reason. $\endgroup$ – March Ho Aug 6 '16 at 15:01
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    $\begingroup$ @March Ho: indeed. I complained to Nature, and they told me that they cannot police that and can only rely on the honor of their authors. Then I complained to Professor John Savill, the CEO of the Medical Research Council, who wrote a fiery letter to the scientist in question - and he still refused to comply. But then eventually Doolan & Colby came to the rescue. Molecular biology against bullies: isn't that great? :) $\endgroup$ – aag Aug 6 '16 at 17:28

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