Hamster hybridoma antibody cloning and antibody sequencing is the most challenging among hybridoma antibody cloning and sequencing because sparse sequence and structural information for hamster antibodies is available in public sources. Back to 2011 when Syd Labs started to provide the services of hamaster hybridoma antibody cloning and antibody sequencing, one could find in Genbank one partial hamster IgM heavy chain DNA sequence and three hamster IgG DNA sequences only and for kappa light chains only. When we searched published hamster antibody sequences in 2013, we only found a handful. Availability of constant region sequences of a large number of heavy chain and light chains is important for designing PCR primers for hybridoma antibody cloning. However, constant region sequences of three hamster heavy chains (CH) and one kappa and two lambda chains (CL) only have been published by 2013.

Hamsters are widely used to generate monoclonal antibodies against mouse and rat antigens in addition to human antigens. Because of relatively far evolutionary relationship to mice and rats, hamsters generate good immune responses to mouse and rat antigens. Similar to mice and rat spleen cells, hamster spleen cells can also stably fuse with mouse myeloma cells to form hybridoma.

In the past three years, Syd Labs, Inc has successfully cloned and sequenced about half a hundred of hamster antibodies. Because all clients requesting to clone and sequence the hamster hybridoma antibodies are from industry, none of the obtained sequences has been available in public. If the sample quality is good, for example if the hamster hybridoma is well subcloned, our success rate is above 90% with PCR-based hybridoma sequencing. The success rate can reach above 98% with information of N-terminal protein sequence. The success is evaluted by binding affinity test using chimeric antibodies expressed with the obtained sequences. Our advantage of hasmter hybridoma antibody cloning and antibody sequencing results from our optimized general techniques of hybridoma antibody cloning and sequencing and from accumulated hasmter antibody sequences in house. The more hasmter antibodies sequenced, the better our service quality.

References:

1. Cloning of a hamster anti-mouse CD79B antibody sequences and identification of a new hamster immunoglobulin lambda constant IGLC gene region. Haggart R1, Perera J, Huang H. 2013 Immunogenetics. 65(6):473-8.

2. cDNA sequence and Fab crystal structure of HL4E10, a hamster IgG lambda light chain antibody stimulatory for γδ T cells. Verdino P1, Witherden DA, Podshivalova K, Rieder SE, Havran WL, Wilson IA. 2011 PLoS One. 6(5):e19828.