Hi everyone— Thought I might chime in. This technology was developed in my lab based on our studies of toxins and we have methods papers and two patent.
The CPD-tag works great for production of protein with minimal residual residues on the protein. Our studies indicate that it can help solubilize protein during expression. Protein is purified with the C-terminal CPD tag, and the entire CPD and 6xHis-tag are released by addition of InsP6 (phytic acid) which is very inexpensive. The only restriction of the recognition sequence is small-Leu-small so a small residue plus Leucine does remain on your protein. There does also need to be a little flexibility at the end of your protein for access to the protease so our vectors add extra Gly-Ala so the residual is GAAL. CPD is particularly useful for small proteins if the additional residues do not interfere with your assays. https://pubmed.ncbi.nlm.nih.gov/28056928/ https://pubmed.ncbi.nlm.nih.gov/31773580/ https://patents.google.com/patent/US8257946B2/en https://patents.google.com/patent/US8383400B2/en Here is a link to the original paper on mechanism of action of CPD https://pubmed.ncbi.nlm.nih.gov/19620709/ These are not commercially available as despite multiple marketing attempts, there was little interest mid-2010s from biotech for licensing new cloning technologies and we abandoned advanced development If this technology is interesting to you to add to your commercial biotech portfolio, please feel free to reach out to me. Flavio, we can share our vector, but my institution does require an MTA as technology is patented. Most people just use synthetic DNA to generate the exact clone they want so we rarely get requests but happy to share sequences if you need for your synthetic design. Karla Satchell From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> on behalf of Srivastava, Dhiraj <0000bda2be8a675a-dmarc-requ...@jiscmail.ac.uk> Date: Friday, February 16, 2024 at 6:42 AM To: CCP4BB@JISCMAIL.AC.UK <CCP4BB@JISCMAIL.AC.UK> Subject: Re: [ccp4bb] [External] [ccp4bb] Off topic : plasmid expression vector for E.Coli with a cleavable his-tag at the c-ter Hi Flavio While i am not aware of any commercial vector with cleavable c terminal tag, you can easily make your own by introducing protease cleavage site of your choice. However this strategy will results in quite a few extra residues from protease site. An alternative, which is not commercial but available through addgene, is cpd-his tag. It’s a bigger tag (~25 kda) but for me, it helped in solubility and expression. https://www.addgene.org/38251/<https://urldefense.com/v3/__https:/www.addgene.org/38251/__;!!Dq0X2DkFhyF93HkjWTBQKhk!X4zQW9pYePXhpOWDRmDhx8F9CS1fcLsydhbQbLz2By2Pl6vW6Rnzq4kJp8HcdKp1EvVk5oIaXe__iBvZYFJul3k-1u1NgbhjZdL2xUs9Cg$> There are other variants of this vector available on addgene that you can choose from. Depending on your c terminal sequence, it may leave either no extra residues or only one or two residues. Dhiraj ________________________________ From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> on behalf of Flavio Di Pisa <dipi...@gmail.com> Sent: Friday, February 16, 2024 3:33 AM To: CCP4BB@JISCMAIL.AC.UK <CCP4BB@JISCMAIL.AC.UK> Subject: [External] [ccp4bb] Off topic : plasmid expression vector for E.Coli with a cleavable his-tag at the c-ter Dear community, I'm looking for a commercial expression vector for E.Coli with a cleavable his-tag at the c-ter. Anyone can help me? Thank you in advance, Flavio. ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1<https://urldefense.com/v3/__https:/www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1__;!!Dq0X2DkFhyF93HkjWTBQKhk!X4zQW9pYePXhpOWDRmDhx8F9CS1fcLsydhbQbLz2By2Pl6vW6Rnzq4kJp8HcdKp1EvVk5oIaXe__iBvZYFJul3k-1u1NgbhjZdKw_JbGJA$> This message was issued to members of www.jiscmail.ac.uk/CCP4BB<https://urldefense.com/v3/__http:/www.jiscmail.ac.uk/CCP4BB__;!!Dq0X2DkFhyF93HkjWTBQKhk!X4zQW9pYePXhpOWDRmDhx8F9CS1fcLsydhbQbLz2By2Pl6vW6Rnzq4kJp8HcdKp1EvVk5oIaXe__iBvZYFJul3k-1u1NgbhjZdI_8SfJ9g$>, a mailing list hosted by www.jiscmail.ac.uk<https://urldefense.com/v3/__http:/www.jiscmail.ac.uk__;!!Dq0X2DkFhyF93HkjWTBQKhk!X4zQW9pYePXhpOWDRmDhx8F9CS1fcLsydhbQbLz2By2Pl6vW6Rnzq4kJp8HcdKp1EvVk5oIaXe__iBvZYFJul3k-1u1NgbhjZdKP34UlqA$>, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/<https://urldefense.com/v3/__https:/www.jiscmail.ac.uk/policyandsecurity/__;!!Dq0X2DkFhyF93HkjWTBQKhk!X4zQW9pYePXhpOWDRmDhx8F9CS1fcLsydhbQbLz2By2Pl6vW6Rnzq4kJp8HcdKp1EvVk5oIaXe__iBvZYFJul3k-1u1NgbhjZdITNo6mGQ$> ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1<https://urldefense.com/v3/__https:/www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1__;!!Dq0X2DkFhyF93HkjWTBQKhk!X4zQW9pYePXhpOWDRmDhx8F9CS1fcLsydhbQbLz2By2Pl6vW6Rnzq4kJp8HcdKp1EvVk5oIaXe__iBvZYFJul3k-1u1NgbhjZdKw_JbGJA$> ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
