AxioMx’s recombinant antibody services and product development are based on our four core improvements to traditional antibody phage display methodologies. These are:
- Phage library design
- Two novel phage library screening methods: Rapid Liquid Screening & Emulsion Screening
- AxioMx Mutagenesis
Phage Display Libraries
AxioMx has developed human-framework, synthetic recombinant scFv antibody libraries with greater than ten billion (> 1010) unique clones. Our libraries are M13 phagemid libraries. Diversity in our libraries is restricted to the most variable positions in the complementarity determining regions (CDRs) of natural antibody sequences. An AxioMx proprietary method limits the amount of parental clones in the library to less than 1%, making AxioMx’s libraries comparable other phagemid antibody libraries of 1011 clones or larger.
Smaller libraries reduce development costs, making it feasible to develop a range of specialized libraries optimized by framework or binding preference to specific antigen types.
AxioMx’s antibody framework has been optimized for protein expression. Our libraries efficiently generate binders to peptides, protein fragments, full-length proteins, and many small molecules.
Rapid Liquid Screening
Rapid Liquid Screening combines all-liquid biopanning and automation. This approach is amenable to high-throughput production thereby allowing parallel screening of antibody diversity to identify antibodies with the desired affinity and selectivity.
Rapid Liquid Screening is the primary biopanning methodology AxioMx applies to screen both our naive phage libraries and affinity matured libraries. It is very fast and easily scalable.
Micro-emulsion screening is well-suited for discovery of a greater breadth of unique scFv antibodies. AxioMx often applies the approach to customers’ projects where initial binding affinity is less critical than finding a diverse range of binders to the target antigen.
E. coli infected with a library of phage M13 encoding scFv variants are compartmentalized with antigen-coated beads in a water-in-perfluorocarbon oil emulsion. This stable emulsion is the equivalent of 1 x 107 wells. Within each emulsion droplet, multiple copies of the recombinant phage are produced and can bind to the antigen-coated bead. The emulsion is broken and the microbeads are washed and incubated with anti-M13 antibody coupled to fluorescein (FITC). The FITC-labeled beads (with bound phage) are enriched by FACS.
Our proprietary AxioMx Mutagenesis technology mimics our bodies’ affinity maturation process and generates additional mutations in a lower affinity antibody to create a library with good mutational diversity. AxioMx Mutagenesis is very efficient and improves antibody affinities by 4 to 10-fold, or more.
We utilize AxioMx Mutagenesis in custom antibody development as well as affinity improvement of customer existing antibodies. AxioMx is also expert at both error-PCR and Kunkel mutagenesis.