ImmunoProtect™: Hybridoma Sequencing

Your monoclonal antibody (mAb) is extremely valuable, but hybridoma cell lines can become unstable over time and can stop secreting antibody. With ImmunoProtect™, the DNA sequence of your antibody will be determined and can be used to express your mAb recombinantly, should your hybridoma cell line become unstable.

ImmunoPrecise offers the following antibody optimization services:

Antibody chimerization

Our antibody sequencing, cloning, reformatting, and expression service can clone and express any variable antibody domain into any immunoglobulin context required, such as full size mouse, rabbit, chicken, and human format; or chimeras with any Fc region of choice, including effector mutants or Fc fusions.

Antibody humanization

Animal-derived monoclonal antibodies can elicit human anti-animal antibody responses in patients. Therefore it is usually necessary to replace the animal content with the corresponding human amino acid residues. We provide a robust and efficient antibody humanization service, which consistently retains affinity and specificity levels. Our royalty free approach is based on state of the art in silico antibody modeling to identify essential framework and CDR residues for grafting onto a human antibody framework.

Antibody affinity maturation

Antibody affinity is important in therapeutic and diagnostic applications. ImmunoPrecise’s antibody affinity maturation service can improve antibodies affinities by several fold. We can apply different strategies to increase the affinity of your antibody, including gene shuffling and random mutagenesis.

Project Phases Timeframe
I. DNA Sequencing

  • Isotype/subisotype determination
  • Mycoplasma Testing
  • RNA Isolation, cDNA preparation, PCR amplification
  • DNA sequencing of antibody heavy and light chain variable regions

Phase I Deliverables

  • DNA Sequence of antibody heavy and light chain variable regions
  • Sequence Analysis Report
  • Mycoplasma Non-Contamination Certificate

2 Weeks
II. Recombinant Antibody Binding Confirmation and Test Expression

  • Cloning of heavy and light chain variable regions into mammalian expression system
  • Transient test expression of antibody in mammalian cell line to determine expression levels
  • Confirmation of antigen binding by indirect ELISA

Phase II Deliverables

  • 40mL of tissue culture supernatant containing expressed antibody
  • Report on antigen binding and expression level of the antibody

3-4 Weeks

As with all our services, we will tailor an antibody sequencing package for your specific needs. Contact us to discuss your hybridoma health, stability or antibody secretion challenges.