Antibody-Extractor Suite

Hotspot was designed for mining immune repertoire for mutations hotspots in V-Region. Discovering the amino acid being mutated during the hotspots somatic hypermutation process is a valuable starting point in antibody engineering.


Hotspot residues, most likely play a significant role in affinity maturation. Wu-Kabat variability coefficient is a well-established descriptor for AA-position variability. The identification of hotspots is a crucial step in the in-vitro maturation process.

Hotspot extends the Wu-Kabat variability formula and generates an interactive Wu-Kabat variability AA and codon plots. AA and codon position-frequencies charts are plotted on the fly for every selected AA or codon.

Hotspot filters and mines the immune repertoire sequences for specific AA or codon position. The mined sequences can be synthesized and tested or used a starting template in antibody engineering.


Scenario 1

Finds AA or Codon hotspots in VH protein or DNA sequences.

Starting with DNA VH sequences hotspot computes amino acid and codon Wu-Kabat variability. Variability is reported as charts, and the raw data is exported in Excel format for further processing.

Scenario 2

Antibody engineering team wants to fix a ligand into a histidine in VH CDR3.

From 1 million of VH sequences, hotspot computes AA variability and displays the frequency of VH with a histidine in CDR3. Hotspot filter extracts the sequences with histidine in VH CDR3 and exports the sequences in different formats.