Two mainstream strategies have been used to sequence antibody repertoires. The classical strategy is to design primers to cover as many functional V-genes as possible and create a diverse antibody libraries. The modern strategy is to use high-throughput next generation sequencing (NGS) to “deeply” sequencing of binding populations. The abundance of antibody mRNA can also be analyzed in the same time. High-throughput antibody repertoire sequencing protocols from public literatures:

– Degenerate primer design to clone the human repertoire of immunoglobulin heavy chain variable regions.
Identification of the most highly conserved region in framework 1 and framework 4 using iCODEHOP. A set of degenerated 5′ primers for the V(H) genes from human peripheral blood mononuclear cells.

– V-gene amplification revisited – An optimised procedure for amplification of rearranged human antibody genes of different isotypes.
Near to 100% of all functional and putatively functional V-genes in VBASE2 successfully used to amplify rearranged antibody genes of different isotypes.

– MAD-DPD: designing highly degenerate primers with maximum amplification specificity.
A new set of primers for amplification of antibody variable fragments from mouse spleen cells, which theoretically covers very diverse antibody sequences.

The technique of high-throughput sequencing of the antibody repertoire is attractive but challenging for most scientists in academia and industry. Experienced scientists at Syd Labs provide high-throughput antibody repertoire sequencing services if the project is difficult for you.

Disclaimer:
We index the protocols for your information only. It does not necessarily mean that we agree with all of them. You rather than Syd Labs takes full responsibility for using any information described here.