CLICK BIO

SPLITTERA Platform

Why SPLITTERA? CLICK BIO

Based on its specific reaction, SPLITTERA is a simple, fast, efficient and cost-effective universal tool for CLICK-BIO, with multiple applications. It creates a new covalent bond between the protein of interest and the payload, in vitro or in vivo without modifying the original protein sequence or properties.

SOME USES

  • Pharma product:
    • ADC (Antibody Drug Conjugate)
    • Companion Ab
    • Therapeutic fusion proteins
  • Immuno assays
  • Assay development
  • Screenings

One of the main applications is customization of antibodies, allowing the creation of antibodies libraries functionalized on demand, a technology of great value for antibody-based clinical programs in immunotherapy.

Key Advantages

  • VERSATILITY:
    • Any recombinant protein: No limit in type, size or structure.
    • Any Payload: cytotoxin, tag, fluorescent probe, peptide, DNA, chemical compound, Ab portions…
    • Family of 4 orthogonal split-inteins: Don’t cross react, thus one single protein can be specifically modified with more than one different payload.
  • Split-inteins are not present in the final product (scar of max 6 amino acids)
    • No immunogenic, suitable for in-vivo applications.
    • Conjugated proteins show the same properties as the unconjugated (antigen specificity, internalization)
  • Intein fusions can be produced in any expression host, without affecting expression and solubility.

Properties

  • 4 different orthogonal pairs of inteins.
  • N-ints: 80aa, C-Ints: 40 aa.
  • Extremely FAST, high reaction YIELD.
  • 100% SPECIFICITY, no interaction with other split inteins
  • Minimal amino acid requirement.
  • Active under a wide range of conditions (pH 6-9 and 4-45°C).
  • Accepting detergents and chaotropic agents
Figure 2. Extremely fast ligation reaction reaching completion after only 30 minutes

Multilabelling

4 orthogonal split-inteins to incorporate different payloads on the same protein, conferring extra added value.

Bioconjugates: site-specific modification of proteins

Conjugation of any protein (Ab, Fabs, ScFv, nanobodies, bispecifics,etc.) with the desired payload (toxin, fluorescent or colorimetric probeschemical compound, DNA, peptides, proteins, IgG derivates) to generate the final product for a given application.

  • Homogeneous, site-specifically modified protein
  • No modification or mutation on functional domain of mAb
  • Controllable DAR (Drug to Antibody Ratio) 2, 4, 2+2, different payloads
  • No need for non-natural amino acids or sugars, enzymes or cofactors

Bispecifics: Ab formats and Fc-Fusions

The orthogonality of Splittera split-inteins allows different strategies to obtain bispecifics, other IgG formats as well as Fc-Fusions.

Using a different pair of Splittera split-inteins, these molecules can also be labelled with any payload.

In vivo and in vitro

SPLITTERA split inteins are not present in the final product, leaving a scar of maximum 6 aminoacids. 

Consequently, conjugated products show the same properties (antigen specificity, internalization, etc) as the original, and are not immunogenic, making it possible to use SPLITTERA for in in-vivo as well as in in-vitro applications.

Figure 3. Antigen specificity comparison between a non-modified IgG (mAb, blue line) and the same IgG with 2 toxin molecules fused to the HC (mAb-tox, green line)