Active Motif offers custom antibody conversion services for customers that have multiple hybridoma clones that they would like to convert to an AbFlex® recombinant antibody.
AbFlex® antibodies are recombinant antibodies derived from mouse, rat, or rabbit monoclonal antibodies. In addition to flexible labeling and purification options, our AbFlex® recombinant antibodies allow for sustainable production from transient transfection of a DNA plasmid encoding the antibody heavy and light chains.
For more information about AbFlex® antibodies, please visit the AbFlex® Recombinant Antibody landing page.
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Hybridoma Conversion Service includes:
Customers submit Hybridoma clones or total RNA purified from hybridoma cultures, we then proceed with the following steps:
- Milestone 1: Hybridoma NGS.
RT-PCR is performed on mouse or rat IgG Heavy and Light chain genes, followed by NGS of the IgG derived transcripts. Transcripts are then ranked by abundance and high scoring heavy and light chain sequences can be chosen for subsequent cloning.
- Milestone 2: Custom Cloning.
Customers can choose to have the appropriate heavy and light chain CDRs of their antibody of interest cloned into a standard IgG backbone vector, or, for an additional cost, cloned into the proprietary AbFlex® mouse IgG2a expression vector, which includes sortase A5, avidin, and 6X-His tags.
- Milestone 3: Expression & Purificaton.
Clones for Heavy & Light chains are sequence verified and pilot expression transfection experiments are performed to determine the yield of the resulting full-length antibodies. Customers are also provided with purified antibody from the pilot phase to verify the correct binding Heavy & Light chains were chosen for cloning.
- Milestone 4: Antibody Production.
The customer chooses the amount of purified antibody they would like to have produced, the transfections are scaled appropriately, and purified antibody is delivered, in the desired quantity chosen by scale.
- Milestone 5: Antibody Conjugation.
For customers that have chosen our AbFlex® backbone vector, custom conjugation services are also offered at this step (Sortase A5 assisted enzymatic conjugation).