Precision Capturing

Conventional multi-step chromatography procedures in downstream bioprocessing impacts, more than any other manufacturing step, the overall manufacturing costs, process development and manufacturing timelines, and manufacturability risks for biopharmaceuticals.
Navigo Proteins’ Precision Capturing platform offers a more efficient alternative, based on a single affinity chromatography step. Precision Capturing affinity chromatography dramatically simplifies the downstream procedure, strongly reduces number of process steps, effectively shortens development and manufacturing timelines, and in consequence enables robust, scalable and economic industrial manufacturing processes.

This way, notably in late stage and for market supply Precision Capturing affinity chromatography significantly reduces time, risk and COGS.

Navigo Proteins envisions affinity chromatography as a generic platform process for any, even complex biopharmaceutical product. We provide solutions for all biological product classes, including antibodies, antibody fragments, fusion proteins, enzymes, hormones, blood factors, as well as vaccines, viral vectors, virus-like particles, or even entire cells.

We discover and develop affinity ligands with best-in-class binding capacity and selectivity, as well as chemical and caustic stability. Our affinity chromatography ligands are fully patent protected to our customers.

Navigo Proteins co-operates with renowned partners for the GMP-compliant coupling of our ligands to supply regulatory well-accepted resin or membrane-based ready-to-use chromatography matrices.

ANTIBODY PURIFICATION

The manufacturing of therapeutic antibodies faces ever-increasing costs and regulatory pressure in an increasingly competitive environment. In 2013 more than 8 metric tons of antibodies have been manufactured. The antibody purification market segment grows with an annual CAGR of 8 - 12% and has reached a volume far beyond 1 billion US dollars annually. Even though the generic antibody downstream platform process by Protein A affinity is firmly established, the manufacturing industry expresses a clear demand for more effective and cheaper manufacturing procedure.

Navigo Proteins provides a solution for all these requirements by having developed innovative, proprietary next generation Protein A affinity ligands are designed to meet these requirements. Our Protein A ligands have both higher caustic stability and dynamic binding capacity, compared to even commercially available gold standard market products.


Navigo’s Protein A variants have superior dynamic binding (DCB) capacities

Navigo’s Protein A variants show superior caustic stability

Elution profile: pH optimum of Navigo’s Protein A variants can be precisely engineered

NON-ANTIBODY PURIFICATION

Based on our well-established, robust and unique Affilin® scaffold proteins platform we generate high-affinity protein ligands also for any non-antibody molecules, such as recombinant protein therapeutics, hormones or vaccines, but also for viral vectors, VLP’s, or even entire cells.
Affilin® ligands offer fully adjustable specificities, affinities and pH elution profiles. The selection and screening processes delivers protein ligands specific for any given target, yet literally without any off-target activity. Our Affilin® ligands possess high caustic and chemical stability to cope with CIP and sanitization procedures under harsh conditions. Moreover, and in contrast to antibodies, Affilin® ligands are very small (~8kDa) and stable proteins, but they are broadly engineerable, including easy multimerization for improved capacity, flexible linker design and site-specific, 3D-oriented chemistry. Finally, our Affilin® ligands are amenable to economical production in E.coli at industrial scale.

Navigo Non-Antibody Affinity Ligands have extraordinarily stability that copes with industrial routines and expectations

Navigo Precision Capturing affinity ligands are engineered to match routine caustic DSP processes

Navigo affinity ligands demonstrate minimal capacity loss multi-cycle performance
Performance of Navigo affinity ligands in non-antibody purification processes perform comparable to Protein A gold standard in HCP clearance

​TECHNOLOGY

Our long-standing expertise in protein engineering is at the heart of our Affilin® protein ligands platform. We have implemented and continuously optimized over many years a powerful scaffold technology platform, which yields high-quality binder proteins against literally any target structure, for multiple purposes. One major focus for us is the development of Precision Capturing affinity chromatography ligands for biologics.

Starting point is a selected scaffold protein with favorable characteristics such as:
• small size (<10kDa)
• very high thermal, proteolytic and caustic stability
• no post-translational modifications
• no cysteines
• cost effective production in E.coli

Libraries
High quality and diverse binder libraries are generated at the genetic level from the starting protein scaffold by randomization of a number of surface-exposed amino acid residues. Each randomized position comprises all possible natural amino acids (except Cysteine) which leads to large combinatorial libraries with complexities from 1010 to 1012 variants, depending on the specific display format used.

Selection and Screening
For the subsequent selection process, both phage display and ribosome display are available. The following automated, robotics-assisted high-throughput screening (HTS) key technology identifies Affilin® ligand-enriched selection pools. Further rigorous screening rounds finally yield those Precision Capturing ligands with the desired properties and suitable for large-scale industrial use.
To do so, Navigo Proteins has tailored an HTS platform which allows a flexible assay design and selection, based on multiple parameters. Rapid screening of well beyond 10.000 Affilin® binders per day identifies a set of diverse Precision Capturing candidates from which we select the most promising leads for further development.

High-throughput Screening at Navigo comprises the entire hit-selection and identification process, maximizing throughput and clone diversity


Automated clone-picking

State-of-the-art screening-robotics for characterization of well beyond10.000 Affilin® protein variants per day

In-house developed software for tracking of hits and results throughout the entire development process

State-of-the-art protein purification and analytics is key for seamless ligand identification process

BIOINFORMATICS

In-house developed, customized software supports all processes from early cloning of libraries through screening and data storage, up to analysis of thousands of binder protein variants. Individual software programs address the specific needs of molecular biology, sequence analysis or variants tracking throughout the entire high-throughput screening process, as well as monitoring project work packages and flow.

INTELLECTUAL PROPERTY

Navigo Proteins pursues a stringent policy of patenting its proprietary products and technology. With over 100 patents granted and dozens of applications pending, our assets are secured in all major markets. Moreover, this provides our clients with a safe, undisputed IP protection and thus competitiveness and long-term planning security for any licensed Navigo products and technology.

Format development

The development of optimized Precision Capturing ligand molecules often involves additional chemical or genetic modification to modulate or extend their functionality. For example, multimerization with varying linker lengths and composition may be needed. In addition, site-directed, spatially oriented ligand coupling to the matrix often improves capacity and chromatography performance but requires the introduction of specific amino acids at defined positions. Furthermore, different coupling chemistry may be necessary, depending on the reactive groups of the resin or the membrane support. Navigo Proteins’ scientists can cope with all these continuously differing requirements however, and have gained extensive protein engineering expertise from many projects over the years, to combine all these features into optimized Precision Capturing Affilin® ligands.

Functional characterization

A thorough functional characterization of Precision Capturing ligands from the selection process is absolutely crucial to identify the most promising lead candidates for further development. This comprises a range of biochemical, biophysical as well as functional protein characterization steps in house. In particular, chemical and caustic stability (in static and cyclic performance) are important criteria. Another central criterion is binding capacity (i.e. DBC10, SBC) of the final Precision Capturing ligand-coupled matrix and its process performance in step yields, elution pH and host-cell protein or DNA reduction.


A comprehensive characterization of Precision Capturing ligands is key, including e.g. binding analysis by Biacore or Octet measurements

Classical protein analytical methods; SDS-PAGE

State-of-the-art protein purification procedures at different scales for in-house requirements as well as for clients’ external analysis

In-house developed bioinformatics tools for comprehensive tracking of hits and results

PARTNERING

Biopharmaceutical manufacturing e.g. in antibody production and non-antibody/ vaccine manufacturing is steadily and strongly growing. As a specialist in the field we realize that our partners expect ready-to-use devices guaranteeing the best possible outcome in the shortest possible time. With our clients we precisely define the required specifications related to an individual target of interest and devise a project plan that clearly describes all work packages from target availability, to selection, screening, coupling, and up to the ligands purification performance. In house we conduct the discovery of suitable affinity ligands, carefully select, analyze and supply the best candidates for validation, and carry out further modification/ optimization, as needed. For scaling up and commercial applications that meet all regulatory CMC requirements we co-operate with renowned partners to provide the GMP-compliant, final, ready-to-use affinity ligand-coupled resin or membrane-based affinity matrix. The final result for our partners is a commercial license and the corresponding robust, rapid and reproducible, economic and ready-to-use, turn-key downstream procedure to guarantee commercially competitive purification processes for even the most complex biologics. Navigo Proteins selects strategic partners and specialists with a strong existing stake in the core markets of “biopharmaceutical manufacturing” and ”affinity chromatography” for further development and commercialization of our self-created affinity ligands.