Re: [ccp4bb] anti-hexahistidine antibody
Hi Tomas, Thanks. I think this one is directed at a C-terminal his6-tag. I'm not sure it'll work with an N-terminal tag. Elias From: Tomas Malinauskas <tomas.malinaus...@gmail.com> Sent: Sunday, January 8, 2017 5:39:59 PM To: Fernandez, Elias J Cc: CCP4BB@jiscmail.ac.uk Subject: Re: [ccp4bb] anti-hexahistidine antibody Dear Fernandez et al., maybe this anti-His-tag antibody could be useful: PDB ID 1KTR, its sequence is in the paper https://www.ncbi.nlm.nih.gov/pubmed/?term=12054774 Hope that helps, Tomas On Sun, Jan 8, 2017 at 9:24 PM, Fernandez, Elias J <efern...@utk.edu> wrote: > Dear All, > > Is anyone aware of a recombinant system that produces an N-terminal, > hexa-histidine-directed Fv antibody fragment that can be used for structural > studies (crystallography, SAXS, cryo-EM)? > > Best regards, > > Elias > > >
Re: [ccp4bb] anti-hexahistidine antibody
Dear Fernandez et al., maybe this anti-His-tag antibody could be useful: PDB ID 1KTR, its sequence is in the paper https://www.ncbi.nlm.nih.gov/pubmed/?term=12054774 Hope that helps, Tomas On Sun, Jan 8, 2017 at 9:24 PM, Fernandez, Elias Jwrote: > Dear All, > > Is anyone aware of a recombinant system that produces an N-terminal, > hexa-histidine-directed Fv antibody fragment that can be used for structural > studies (crystallography, SAXS, cryo-EM)? > > Best regards, > > Elias > > >
Re: [ccp4bb] anti-hexahistidine antibody
Besides my private reply, a few suggestions that may be relevant to the broader audience: 1) for the purposes of this email I will pretend that relevant patents do not matter (of course they do but it's too complex to get into details) 2) there are quite a few antibodies out there directed against His(n) sequences. To the best of my knowledge sequences of said antibodies are typically not published because the manufacturers wish to protect their ability to make (and sell) antibodies at a considerable mark-up :) 3) in theory, if someone wanted to make their own anti-His or anti-Flag etc. antibody or Fv they would have to either (a) raise their own in animals/select their own from libraries or (b) take a commercial sample and do a peptide digest + MS(n) study of it in order to determine sequences of the hypervariable regions. Please see my point (1) if you have questions about the legality/ethics of such 'reverse engineering' efforts. I would certainly choose (a), or write to the manufacturer and ask for permission. Once you know the sequence of the mAb that you need to make, there are plenty of options to express either the whole thing or the Fv. We certainly do this kind of work for our clients :) Available options range from E.coli to yeast to HEK/CHO cells. Sky's the limit! Interesting side note: there is a (single!) structure of an anti-FLAG Fab in the PDB database. It has "A" residues where the authors could not reliably place specific amino acid side chains, because the sequence of the original mAb was not available :) Presumably Sigma did not want to share the sequence? https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2242885/ Artem www.harkerbio.com "Will breed miniature camels for antibody purposes..." - Cosmic Cats approve of this message On Sun, Jan 8, 2017 at 4:24 PM, Fernandez, Elias Jwrote: > Dear All, > > Is anyone aware of a recombinant system that produces an N-terminal, > hexa-histidine-directed Fv antibody fragment that can be used for > structural studies (crystallography, SAXS, cryo-EM)? > > Best regards, > > Elias > > > >