Ligand Discovery Projects

July 14, 2017 Andy Davies
Our goal is to provide you with a fully validated bespoke chromatography adsorbent and all the associated process conditions for purification of your target biomolecule.

With increased challenges in downstream purification of biomolecules, standard adsorbents often cannot achieve the purity required. At Astrea Bioseparations Ltd. ('Astrea') we have over 30 years of experience in developing custom affinity ligands and adsorbents that can help achieve safe and effective biotherapeutics. This article describes the various phases needed for the development of a custom affinity adsorbent suitable for GMP manufacture.

 

Ligand Discovery

Assay Development:

Ligand Discovery Projects commence with the development and implementation of high throughput assays for quantification of target biomolecule and impurities to be removed. Typical assays developed include ELISA, SDS-PAGE, HPLC, total protein and functional activity assays.

Ligand Design:

Option 1 - Known receptor structure: Using rational design, molecular docking and virtual
screening to help identify possible sites for ligand binding and potential ligand structures.

Option 2 - Known ligand structure: Ligand selection from Astrea’s Chemical Combinatorial
Library CCL® containing over 100,000 triazine based compounds to provide a focused library for initial screening Focused design based on ligands showing early “activity” against a target.

Option 3 - No prior knowledge of receptor or binding ligands: Selection of binding ligands by screening general ligand libraries comprising diverse ligand structures distributed across a broad region of chemical space.

Ligand Synthesis:

Adsorbents synthesised in library format (PuraPlate™ 96 column block) with ligand structures identified from initial design/virtual screening studies.

Ligand Screening:

High throughput evaluation of existing general ligand libraries and newly designed libraries. Non-bound, wash, elution and CIP procedures investigated.

Verification Chromatography:

Scale-up synthesis and verification chromatography of lead candidates (identified during library screening). Performed using automated chromatography workstations and packed columns with initial investigation of non-bound, wash, elution and CIP steps.

Adsorbent Development

Includes evaluation of:

  • Base matrices;
  • Ligand synthesis method;
  • Immobilised ligand densities;
  • Attachment chemistry (e.g. spacer arms).

Optimisation of adsorbent manufacture and production of written methods in readiness for transfer to Astrea’s manufacturing facility.

Chromatography Development

Optimisation of process conditions (i.e. load, wash, elution and CIP steps) to utilise the adsorbent in the most efficient way and maximise product purity and yield. Performed in parallel to Adsorbent Development An adsorbent cleaning and re-use study may be
performed as part of this work.

Tech Transfer & Validation

Procedures for manufacture of the custom adsorbent will be transferred to Astrea's manufacturing facility situated on the Isle of Man, British Isles. Trial lot(s) are usually made at the 1 litre scale prior to validation of the manufacturing process by production of three conformance batches of custom adsorbent Manufactured batches will be tested for their ability to achieve the process performance defined specifications Samples of the batches of custom adsorbent produced during the validation process will be supplied to the end user
A validation plan and summary report will be produced as a part of this activity.

Regulatory Support

A comprehensive support package of information will be developed and supplied to the end user to assist registration of the purification process with regulatory authorities. The regulatory package will include information derived from the following studies (additional testing and documentation can be included on request):

a) LIGAND LEAKAGE STUDY - Including a forced degradation study under extreme conditions.

b) STABILITY TRIALS - Under a range of normal and extreme conditions, up to one year duration.

c) TOXICOLOGY STUDIES - MTD in vivo and Standard WHO AMES tests.

 

To find out more contact us at [email protected]

All trademarks, trade names, trade dress, product names and logos appearing on this website are the property of Astrea Bioseparations Ltd.

About the Author

Andy Davies

An experienced Sales Director with over 30 years’ experience in the biotech and life sciences sectors, specialising in bioprocessing and bioseparation techniques with degree level Biochemistry from the University of West London. Having started my career working in a research and development laboratory, I have progressed into consultative sales roles for capital high-tech equipment and strategic account management, leveraging my lab experience for large pharmaceutical companies such as GE. I have been with Astrea Bioseparations ('Astrea') since September 2013 with responsibilities covering the global sales and marketing of its products and services. Since joining Astrea, I have been instrumental in developing the product pipeline by successfully launching the ever-growing Evolve and EvolveD column ranges into Astrea’s offering

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