Bioseparation by affinity chromatography

A novel affinity ligand discovery platform

Hundreds of therapeutic proteins are currently in clinical development and on the market. These complex, heterogeneous protein-based products are bio-reaction products that are purified using multiple filtration and chromatographic steps. Affinity chromatography using protein A is first step of most technology platforms for the purification of some classes of monoclonal antibody. However protein A affinity is not universally applicable to all proteins.

NovAliX offers a unique platform for rapid discovery and development of customized synthetic affinity ligands for cost-effective purification of proteins. The components of this platform are: 1) NovAliX’ proprietary chemical microarray SPR screen; 2) the diverse NovAliB library of immobilized small molecules; and 3) NovAliX’ extensive synthetic-chemical and biophysical services (e.g. Hit progression chemistry and protein crystallography). Potentially useful ligands are characterized against a number of chromatographic performance parameters. The ligands thereby discovered are specific binders, with high chemical stability and submicromolar to lower micromolar affinities; ideal for purification or chromatographic applications. Such small molecule affinity ligands are economical to produce. The discovery process itself establishes the site for attachment of the linker. A functionalization at this given attachment point allows functionalizing the ligands and facilitates immobilization to the preferred chromatographic support.

Workflow

The target protein is screened against the entire NovAliB library via the NovAliX chemical microarray SPR platform. Additionally, a counter-screen with potential feed impurities is performed to verify the required specificity. Hits are selected for immobilization for example on agarose for chromatographic characterization. Miniaturized microtiter plate assays rapidly determine binding behavior, selectivity, isotherms, kinetics and chemical stability of the hit structures. Dynamic binding capacity and preparative column chromatography are investigated. Affinity ligands performance can be further optimized  by employing the NovAliX expertise more commonly used to support drug discovery programs; hit to lead and lead optimization and SBDD.

Features

  • Proprietary screening approach for the discovery of high quality affinity ligands
  • Rapid results – within 6 months for full chromatographic characterization of screening hits
  • Universally applicable to all soluble proteins
  • High specificity for defined binding sites can be developed
  • Development of target specific and group specific (e.g Fab) binders possible
  • Screening of immobilized molecules facilitates transfer to chromatography format
  • Fine-tuning of performance is a clearly defined process
  • Variation of linker for the coupling to the chromatographic support matrix of choice
  • Novel affinity systems are characterized by chromatographic performance
  • Leverages expertise developed for medicinal chemistry and SBDD
  • Evaluation and optimization of the best chemical routes

Benefits

  • High probability of success – applies established paradigms
  • High novelty content – strong intellectual property protection – patentable