High-throughput sequencing and immunology

• • To amplify the VDJ fragment with PCR, primers are designed for each V and each J gene in the genome
  • Blood is extracted and PCR is run on the mixed cells
  • The resulting DNA is sequenced and the identity of each V, D, J determined by blasting against a database of VDJ genes
  • The number of each VDJ combination can be determined as a result
  • The big dream is to help identify the origin of allergies (which antibody? what does it bind to?) with this, to detect diseases before they produce symptoms (e.g. by finding antibodies in them) or to find antibodies against tumors. No one has achieved this yet.

• • Q: Somehow we think that the heavy chains are more important than the light chains, and CDR3 more important than CDR1 and CDR2

• • The heavy chains are encoded by one of the few loci that can recombine somatically, the IGH-locus