Enabling rapid liquid and freeze-dried formulation design for the manufacture and delivery of novel biopharmaceuticals
Lead Research Organisation:
University of Manchester
Department Name: Chem Eng and Analytical Science
Abstract
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Publications
Bye J
(2021)
ATP and Tri-Polyphosphate (TPP) Suppress Protein Aggregate Growth by a Supercharging Mechanism.
in Biomedicines
Bye JW
(2019)
Controlling Phase Separation of Lysozyme with Polyvalent Anions.
in The journal of physical chemistry. B
Corbett D
(2019)
Measuring Nonspecific Protein-Protein Interactions by Dynamic Light Scattering.
in Methods in molecular biology (Clifton, N.J.)
Curtis R
(2019)
Thermodynamics of Phase Equilibria in Food Engineering
Douglas JF
(2017)
Hard Spheres with Purely Repulsive Interactions Have Positive Diffusion Interaction Parameter, kD.
in Biophysical journal
Hebditch M
(2019)
Models for Antibody Behavior in Hydrophobic Interaction Chromatography and in Self-Association.
in Journal of pharmaceutical sciences
Hebditch M
(2017)
Sequence composition predicts immunoglobulin superfamily members that could share the intrinsically disordered properties of antibody CH1 domains.
in Scientific reports
Hebditch M
(2017)
Protein-Sol: a web tool for predicting protein solubility from sequence.
in Bioinformatics (Oxford, England)
Hebditch M
(2019)
Web-based display of protein surface and pH-dependent properties for assessing the developability of biotherapeutics.
in Scientific reports
Hebditch M
(2020)
Modelling of pH-dependence to develop a strategy for stabilising mAbs at acidic steps in production.
in Computational and structural biotechnology journal
| Description | Our novel analytical device has been able to determine and identify complex interactions between protein molecules that are not detectable by standard methods. It is able to identify the relative amounts of two antibodies in the same solution based only on optical spectra. The device is also able to detect aggregate formation with much higher sensitivity than standard SEC detectors, enabling much earlier use of SEC in aggregation kinetics studies. This means that the device is very suitable for monitoring complex degradation pathways in protein formulations. It has attracted wide interest from industry, and is being developed further now in a partnership with Pall Europe. We have revealed complex concentration dependent behaviour in aggregation kinetics of several proteins, in which aggregation slows at higher concentrations contrary to normally expected behaviour. We have been able to determine the mechanisms for this via rheology and other kinetic measurements. A key factor controlling the critical quality attributes (CQAs) of biopharmaceuticals are hot spots or sticky regions on protein surfaces composed of hydrophobic groups. Through an informatics analysis of tyrosine groups in antibody complementarity regions, we have shown that empirical models for calculating the stickiness of hydrophobic surfaces can be improved by incorporating a shape-dependent term. (Hebditch et al (2019) J Pharm Sci 108:1434). Large-scale measurement of biophysical properties for antibodies has opened the way to improved prediction models for CQAs. We have shown, using machine learning applied to sequence, that much of the variation in biophysical properties can be quantified in terms of sequence features such as charge and hydrophobicity. This for instance has allowed us to develop an improved model for predicting hydrophobic interaction chromatography (HIC), which indicates protein charge is a significant factor in controlling the retention time. The finding is directly relevant for formulation development as HIC is frequently used as a high-throughput readout for assessing biopharmaceutical developability (Hebditch M and Warwicker J (2019) PeerJ. DOI:10.7717peerj.8199). We have discovered a novel class of promising excipients based on poly-phosphates for improving the CQAs of a broad range of therapeutics. We have shown their mechanistic action is different from other classes of excipients. The multivalent ionic excipients bind to and supercharge proteins leading to an increase in their colloidal stability (Bye J and Curtis R (2019) J Phys Chem 123: 593). In mixtures with other excipients, they exhibit synergistic effects, which provides much scope for developing formulations with even greater effectiveness at stabilizing proteins and biopharmaceuticals. The project enabled us to collaborate with Oxford Jenner Institute to optimise the stability of ChAdOx vectors used in their Covid vaccine through formulation and freeze drying. The collaboration and technologies developed in the EPSRC project led to further feasibility funding from the EPSRC/DoH VaxHub to develop freeze-dried ChAdOx doses stable at 30-45C, suitable for delivery to LMICs I the absence of a robust cold chain. |
| Exploitation Route | The methods developed will be of very high value to the biopharmaceutical industry by improving both formulation design and also forced degradation for formulation screening. The analytical methods we have developed are now being translated through partnership with Pall Europe, in an EPSRC EngD, to develop it for inline bioprocess monitoring purposes. The formulation data and ML methods for predicting HIC properties from protein sequence will be of immense value to industry in the design of bioprocesses and protein formulations. We have developed our web-based tool (www.protein-sol.manchester.ac.uk), which is publicly available, for predicting protein solubility and aggregation propensity from protein structures (and models), which are key factors in controlling biopharmaceutical development. Key predictive indicators calculated by our server are charge and hydrophobic patches, and the pH-dependence of stability. Our server that includes these tools is currently supporting ~ 300 runs per month from external users (Hebditch and Warwicker (2019) Sci Rep 9:1969) and has already been cited in numerous publications concerned with predicting developability of biopharmaceuticals. We have been awarded industrial acceleration impact funds from the University of Manchester in a collaboration with Albumedix for investigating the potential of using the multivalent ionic excipients in mixtures with recombinant human serum albumin (HSA) to improve upon Albumedix formulation technology. The preliminary experimental studies have demonstrated the effectiveness of the excipients for stabilizing liquid formulations. We are currently exploring patent opportunities for the technology. Along these lines, we are working with the Centre for Molecular Immunology (CIM) in Cuba and Immunocore Ltd to find optimal formulations with the excipients for stabilizing IL-2 based biopharmaceuticals from CIM and ImmTAC bispecific antibodies from Immunocore. Both types of biotherapeutics are already in various stages of clinical trials. Additional collaborations that have arisen based on project outputs include working on vaccine formulations for Zika (Brazil), and Covid (Jenner UK, and TMU Germany) and developing methods for predicting rheological properties of concentrated antibody formulations with Boehringer-Ingelheim (Germany). |
| Sectors | Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
| Description | The early findings on the novel analytics from the joint project between UCL and Manchester has led to a collaboration between UCL and Pall Europe via an EPSRC EngD project. This is also aiming to lead to a commercial partnership in the near future to exploit the technology which has now been filed for a patent. Analytical and formulation techniques are now being explored for their use in the gene therapy sector, via a PhD studentship within the BBSRC ABViP CTP consortium (UCL/Oxford/Oxford Biomedica). The formulation prediction approaches are being exploited through a collaboration with Albumedix, supported by Impact acceleration impact funding in Manchester. Visibility of this project has led to two collaborations on vaccine formulation development. One with Oxford (Jenner) to develop a freeze dried formulation and process for ChAdOx. A second is to develop an undisclosed vaccine formulation with Intituto Butantan in Brazil. Along these lines, we are working with the Centre for Molecular Immunology (CIM) in Cuba and Immunocore Ltd to find optimal formulations with the excipients for stabilizing the IL-2 based biopharmaceuticals from CIM and the ImmTAC bispecific antibodies from Immunocore. Both types of therapeutics are already in various stages of clinical trials. Lastly, the predictive modelling approaches that have been developed through the project work are now being exploited in a collaboration with Boeringher Ingelheim for assessing the critical attributed of concentrated antibody formulations. |
| First Year Of Impact | 2021 |
| Sector | Healthcare,Pharmaceuticals and Medical Biotechnology |
| Impact Types | Economic |
| Description | BBSRC IAA |
| Amount | £8,912 (GBP) |
| Funding ID | BB/S506692/1 |
| Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 10/2019 |
| End | 04/2020 |
| Title | A publicly available database for solubility prediction of biopharmaceuticals |
| Description | A key stumbling block to predicting solubility or aggregation propensity is the lack of comprehensive database relating behaviour to structure along with a suite of tools/algorithms for mining the data and testing predictions. Towards this end, we have developed a publicly accessible web server that uses our in-house and freely available software and algorithms for solubility prediction, in a user-friendly graphics window. The software is used to calculate a variety of different properties from protein amino acid sequence and/or protein structural data. Properties calculated for sequences include: amino acid compositions; combinations of charges to give net predicted charge, fraction charged, predicted isoelectic point, ratio of conservative amino acids; propensity for disorder; and ß-strand/sheet forming propensity. These properties are calculated as averages over the entire sequence and as minimal/maximal values in windows of 21 and 51 amino acids along the sequence. For structures and structural models, properties calculated include: patch non-polarity, calculated as non-polar solvent accessible surface area (SASA) divided by polar SASA, size distribution of charged patches (positive and negative, including the largest patches) from Finite Difference Poisson Boltzmann (FDPB) calculations; average contact order for heavy atoms in the protein, relating to the degree of close packing as an empirical proxy for structural stability. Charge patches are dependent on ionic strength and pH, features that are also part of FDPB calculations, which allows us to account for ionic strength and/or pH effects on solubility. The philosophy of our website is to make all software available both to use online, and as downloadable code for in-house use. |
| Type Of Material | Antibody |
| Year Produced | 2016 |
| Provided To Others? | Yes |
| Impact | none as yet |
| URL | http://www.protein-sol.manchester.ac.uk/ |
| Description | Albumedix |
| Organisation | Albumedix Ltd |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | Through our project, we have developed ways for predicting, and controlling critical attributes for liquid formulations of therapeutic proteins, the most important being the aggregation propensity. Aggregation is strictly regulated and must be prevented due to there being an increased risk of immunogenecity. Often this is accomplished by using low temperature supply chains with additional added cost. An alternative preventive strategy is to add excipients to formulations. We have identified multi-valent ions with superior aggregation suppression effects due to their ability to bind to and over-charge proteins. Albumedix markets a recombinant protein, human serum albumin (rHSA), which itself is used as an excipient for difficult to stabilize peptides. We have shown the feasibility of combining multi-valent ions and albumin in order to create therapeutic protein formulations with superior stabilizing properties. We have created an initial dataset of biophysical measurements on HSA solutions without any API in the presence of multi-valent excipients. The dataset provides a knowledge base to be used in further studies on HSA solutions and opens up additional business opportunities for Albumedix to use HSA in co-formulations with difficult to stabilize therapeutics. The project has been funded through pump-priming funds at the University of Manchester. We are seeking follow-on funding through knowledge transfer partnership (KTP) awards and InnovateUK. |
| Collaborator Contribution | In addition to providing gram quantities of protein material, Albumedix is providing access to state of the art analytical technologies and more importantly has expertise and experience in developing biopharmaceutical products with albumin as an excipient. This is essential for identifying patent opportunities from understanding state of the art in formulation technology, for exploiting their network of industrial partners in future applications, and overseeing initial trials to demonstrate the safety of multi-ionic excipients. The research made possible through the collaboration is also part of a PhD student training and will form the basis of one to two chapters in a dissertation and eventually lead to publication of the work. |
| Impact | No outputs yet. |
| Start Year | 2019 |
| Description | Boehringer-Ingelheim |
| Organisation | Boehringer Ingelheim |
| Country | Germany |
| Sector | Private |
| PI Contribution | We were awarded an EPSRC case studentship with Boehringer-Ingelheim as the industrial partner. The aim of the project is to develop a new method for predicting the properties of concentrated protein solutions using only measurements made on diluted protein solutions. The method we are developing is based on measuring the Huggin's coefficient. The studentship started in April 2022. |
| Collaborator Contribution | The industrial partner has provided £40,000 of funding to the project to cover travel and consumables costs and has also provided £50,000 worth of antibody material including two different antibody formulations in gram quantities. |
| Impact | none so far |
| Start Year | 2022 |
| Description | Immunocore |
| Organisation | Immunocore Ltd |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | We were awarded a BBSRC-DTP case studentship with Immunocore as the industrial partner. The aim of the project is to characterize the reversible self association behaviour of bispecific antibodies manufactured by Immunocore and to understand how the behaviour can be tuned by using multivalent ions as excipients. The studenship started in September 2022. |
| Collaborator Contribution | Immunocore has provided £22000 funding to cover consumables and travel costs and $10000 in kind contribution including protein material. The student will visit Immunocore for training and skill development for a career in industrial research and development of biopharmaceuticals. |
| Impact | none yet |
| Start Year | 2022 |