Hi Yadira,

I am not sure what exactly you want to do with your sequences but for analysis of Ig variable genes, I would recommend  using the IMGT database http://www.imgt.org/IMGT_vquest/vquest?livret=0&Option=humanIg. This is a powerful and free analysis program that contains all known germline Ig gene sequences from a range of organisms. Simply paste in your sequence, and the program will return the V, D and J usage, the number of mutations in each framework and CDR region and a whole load of other information about your sequence.

Thanks for your suggestion Andrew!

I am supposed to get the sequence to produce a recombinant antibody afterwards. I've been using the new database of BLAST, which is the IgBLAST to look for my sequence. What I understand is that my sequence - since it is supposed to be unique- should have less than 100% identity with with the sequences registered in the database, especially in the CDR3. However, I haven't been able to get that. Thus, I've been wondering if I using my sequencing data and parameters in the right way.

hi You can also try joinsolver. my thesis is on this human B cell and i used this online data base which provide wide range of information too.

best of luck

please see papers of  Lipsky group

Hi Yadira,

I just came across this question and thought I’d put an update in, for the benefit of anyone doing similar research today using antibody sequencing.

Antibody Sequencing can now be done with a very high degree of confidence directly from an antibody protein sample and without need to reference IMGT or use igBLAST - the technique involves enzymatic digestion, tandem LC-MS/MS mass spectrometry, post-analysis of the data using de novo peptide sequencing software, and sequence assembly/coverage/confidence assessment.

Here are some details on how the method works: Antibody Sequencing (https://www.rapidnovor.com/antibody/)

Here are some links if you are looking for expert help with this step:

Antibody Sequencing Services (https://www.rapidnovor.com/antibody-sequencing-service/)

Antibody Sequencing plus Expression Services (https://www.rapidnovor.com/recombinant-antibody-expression-service/)

Thank you.

As I understood you have the Peptide Sequence. You can change it by reverse translate (online programm) into DNA then put it into igBlast and see your result.

About de novo sequencing, depending on what programm you are using it is always different, but I can recommend this paper Article PEAKS: Powerful software for peptide de novo sequencing by t...

To a successful outcome!