Regenerative BioCrystallisation
Lead Research Organisation:
Loughborough University
Department Name: Chemical Engineering
Abstract
Advances and breakthroughs in target-oriented biotechnology research are vital to the development of new biopharmaceutical products and treatments. Despite the increasing success in discovering protein-based medicines and progress on upstream bioprocess, manufacturing especially downstream bioseparation process at scale remains a significant challenge in the biopharmaceutical industry at present, such as (a) low through-put production, (ii) costly and complex downstream steps, and (iii) impurity and instability during the storage and transport. The bottleneck in downstream biomanufacturing limit the scaling up in downstream bioprocess, such as bioseparaton, limit the economic scale-up benefit from upstream transform to downstream, and, accordingly, limit the wide application of the advanced biopharmaceuticals to the patients, and limit the disease survival rates, such as cancer. Biocrystallisation is an alternatives to the current complex downstream bioseparation technology, such as multiple chromatography, which are high cost, low efficient and hard to scale up.
The vision here is to develop an interdisciplinary technology (bioengineering, bioseparation and biochemical engineering) de-bottleneck of downstream bioprocessing, integrating upstream (producing protein) and downstream bioprocess (biocrystallisation and bioseparation). Regenerative biocrystallisation leads to a novel and simplified biomanufacturing technology to directly grow biocrystal/protein crystals in vivo. The biocrystallisation inside cells ensure deliver 21st biopharmaceuticals with high purity and quality with low cost, high efficiency and easy to scale up. This technology is transformative, but there are with relatively high risks associated with this adventurous, original and ambitious project. Interdisciplinary experts will help to steer the project. The new scientific understanding in this project and the risks has the very real potential to deliver an new insight technology, and bring new insight into biomanufacturing inside cell.
This research will enable the PI and PDRA to develop interdisciplinary collaborations with existing EPSRC Centres of Excellence, UK biopharmaceutical industrial partners and academic groups internationally.
The vision here is to develop an interdisciplinary technology (bioengineering, bioseparation and biochemical engineering) de-bottleneck of downstream bioprocessing, integrating upstream (producing protein) and downstream bioprocess (biocrystallisation and bioseparation). Regenerative biocrystallisation leads to a novel and simplified biomanufacturing technology to directly grow biocrystal/protein crystals in vivo. The biocrystallisation inside cells ensure deliver 21st biopharmaceuticals with high purity and quality with low cost, high efficiency and easy to scale up. This technology is transformative, but there are with relatively high risks associated with this adventurous, original and ambitious project. Interdisciplinary experts will help to steer the project. The new scientific understanding in this project and the risks has the very real potential to deliver an new insight technology, and bring new insight into biomanufacturing inside cell.
This research will enable the PI and PDRA to develop interdisciplinary collaborations with existing EPSRC Centres of Excellence, UK biopharmaceutical industrial partners and academic groups internationally.
Planned Impact
Impact Summary
The success of the proposed is to develop a novel technology to efficiently deliver 21st century biopharmaceuticals by replacing the current complex chromatographic approaches in downstream manufacturing. The new technology will lead new generation of biotechnology for biopharmaceuticals manufacturing with high efficiency and low cost. In addition, crystalline biopharmaceuticals improve the stability, enabling easy and low-cost storage and delivery than traditional product forms, and reduce the like hood of side-effects and can be expected to increase disease survival rates. A Steering Committee from interdisciplinary research area will steer the project. This proposal will intent to develop collaborations with EPSRC Future Manufacturing Research Hubs and other UK collaborations in interdisciplinary research fields.
This proposal will liaise closely with the technology transfer research office at the university to identify potential opportunities for Intellectual Property arising from the project. It will potential reinforce the research vision by further develop the intellectual properties to apply the new technology on the scaling up of the industrial biopharmaceutical manufacturing for future beneficiaries of UK biopharma industrial and wider biotechnology stakeholders. This research will take activities, such as international conference (ACS and AICHE) and UK conferences, high-quality publication to assure research is well publicised to ensure maximum exposure to the scientific and industrial community; such as outreach, school visit, engagements in University / Departmental Open Days, and science festival to assure research is well publicised to ensure maximum exposure to the general public. Dedicated designed web, together with other online social networking, such as twitter and LinkedIn, will be used to broadcast the new technology developing, public science and sharing the new research ideas, for enhancing the connection between academic and industrial collaborations, and further raise the public engagements with science.
The success of the proposed is to develop a novel technology to efficiently deliver 21st century biopharmaceuticals by replacing the current complex chromatographic approaches in downstream manufacturing. The new technology will lead new generation of biotechnology for biopharmaceuticals manufacturing with high efficiency and low cost. In addition, crystalline biopharmaceuticals improve the stability, enabling easy and low-cost storage and delivery than traditional product forms, and reduce the like hood of side-effects and can be expected to increase disease survival rates. A Steering Committee from interdisciplinary research area will steer the project. This proposal will intent to develop collaborations with EPSRC Future Manufacturing Research Hubs and other UK collaborations in interdisciplinary research fields.
This proposal will liaise closely with the technology transfer research office at the university to identify potential opportunities for Intellectual Property arising from the project. It will potential reinforce the research vision by further develop the intellectual properties to apply the new technology on the scaling up of the industrial biopharmaceutical manufacturing for future beneficiaries of UK biopharma industrial and wider biotechnology stakeholders. This research will take activities, such as international conference (ACS and AICHE) and UK conferences, high-quality publication to assure research is well publicised to ensure maximum exposure to the scientific and industrial community; such as outreach, school visit, engagements in University / Departmental Open Days, and science festival to assure research is well publicised to ensure maximum exposure to the general public. Dedicated designed web, together with other online social networking, such as twitter and LinkedIn, will be used to broadcast the new technology developing, public science and sharing the new research ideas, for enhancing the connection between academic and industrial collaborations, and further raise the public engagements with science.
People |
ORCID iD |
Huaiyu Yang (Principal Investigator) |
Publications
Lin Z
(2023)
Positive and negative effects of graphite flake and monolayer graphene oxide templates on protein crystallization
in Particuology
Sun M
(2022)
Macromolecular protein crystallisation with biotemplate of live cells.
in Scientific reports
Tao T
(2022)
Enhancement of protein crystallization with the application of Taylor vortex and Poly(ionic liquid)s
in Chemical Engineering Science
Tao T
(2023)
Taylor vortex-based protein crystal nucleation enhancement and growth evaluation in batchwise and slug flow crystallizers
in Chemical Engineering Research and Design
Tian W
(2023)
Protein Nucleation and Crystallization Process with Process Analytical Technologies in a Batch Crystallizer.
in Crystal growth & design
Tian W
(2023)
Protein crystallisation with gas microbubbles as soft template in a microfluidic device
in Molecular Systems Design & Engineering
Tian W
(2021)
Protein crystallisation with air bubble templates: case of gas-liquid-solid interfaces
in CrystEngComm
Wang L
(2021)
Wet Milling, Seeding, and Ultrasound in the Optimization of the Oiling-Out Crystallization Process
in Industrial & Engineering Chemistry Research
Wang L
(2021)
Investigation of Agglomeration in the Presence of Oiling Out in the Antisolvent Crystallization Process
in Industrial & Engineering Chemistry Research
Yang H
(2019)
Gravity on Crystallization of Lysozyme: Slower or Faster?
in Crystal Growth & Design
Description | New method/technology: 1. Success in using the gene synthesis method to drive cell manufacture of more target proteins inside the cell. The protein will be produced and with help of the functional tags attached to the protein, it is possible to easily distinguish the target protein with fluorescent colour under a microscope. 2. Success in obtaining protein crystals inside live cells - E Coli cell, with designed DNA plasmid, injected into the cell for the cell culture, the protein crystals have been observed under microscope and SEM. 3. Success in developing stable cell line for producing target protein and target protein crystals. New research questions opened up for what is the mechanism for the cell to control the nucleation or crystal growth inside the cell. New research networks and collaborations were established with Imperial College London, Tianjin University, Xian Jiaotong University, and the Academy of Agricultural Sciences, China. The research will lead to a potential pathway to the commercialization of the technology in biopharmaceutical manufacturing for lower cost and higher efficiency, which will benifit the patients and will contribute to build a healty future. |
Exploitation Route | 1. The heterologous expression method to produce the protein can be used by other researchers, including the design of plasmids, vectors, and sequences. 2. The method of cell culture for recombinant protein crystallization can be used by other researchers, for generating more protein crystals inside cells. 3. The technology of purification of protein inside cells can be developed to be used in the biopharmaceutical industry. 4. The cell lines for producing target protein and target protein crystals can be used in biopharmaceutical industry. |
Sectors | Healthcare,Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology |
Description | AACME PhD studentship on biocrystallisation |
Amount | £72,000 (GBP) |
Organisation | Loughborough University |
Sector | Academic/University |
Country | United Kingdom |
Start | 10/2021 |
End | 09/2024 |
Description | Advanced miscroscope for protein and cells |
Amount | £28,000 (GBP) |
Organisation | Loughborough University |
Sector | Academic/University |
Country | United Kingdom |
Start | 06/2022 |
Description | New Generation Technology of Biopharmaceutical Manufacturing for Therapeutic Proteins |
Amount | £178,000 (GBP) |
Funding ID | 1681 |
Organisation | Loughborough University Enterprises Ltd |
Sector | Private |
Country | United Kingdom |
Start | 10/2022 |
End | 10/2023 |
Title | E Coli cell line to express recombination protein |
Description | We synthesized the gene for Cry protein from Genescript. As this gene was taken from Bt bacteria so the GC content was very high. We performed codon optimization of gene sequence for better expression in the Ecoli system. Further, this cry gene was cloned into a pET28 vector for heterologous expression in E.coli. To further access the protein quality and behavior we induced a fused fluorescent tag with cry protein. |
Type Of Material | Cell line |
Year Produced | 2021 |
Provided To Others? | No |
Impact | This cell line can be used to produce proteins and protein crystals in E. Coli cells, and the proteins can be easily observed under optical and fluorescent microscope. |
Title | Hek cell line to express recombination protein |
Description | We have developed transformation and stable HEK cell line for expression protein and protein crystals inside cells. |
Type Of Material | Cell line |
Year Produced | 2022 |
Provided To Others? | No |
Impact | With the cell lines we developed, it is able to produce multiple different combination proteins and protein crystals. |
Description | Biocrystallisation with cell template |
Organisation | Jilin Agricultural University |
Country | China |
Sector | Academic/University |
PI Contribution | We collaborated on biocrystallisation with the living cells as templates. In our team, we did knowledge exchange and training of protein crystallisation technology. |
Collaborator Contribution | We collaborated on biocrystallisation with the living cells as templates. In the collaborative team, they did the investigations on the influence of living cells in protein crystallisation, including observing the protein crystallisation process and the characterisation of the protein crysttals. |
Impact | We have one collaborative publication. doi.org/10.1038/s41598-022-06999-7. |
Start Year | 2021 |
Description | Biocrystallisation with different cell systems |
Organisation | University of Derby |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We have developed the biocrystallisation technology in E Coli cell systems, we are trying to develop the biocrystallisation technology in other cell systems. In our team, we did the biocrystallisation in mammalian cells and BT cells. |
Collaborator Contribution | We have developed the biocrystallisation technology in E Coli cell systems, we are trying to develop the biocrystallisation technology in other cell systems. In the collaborative team, they have designed the plasmid for cell culture and provided many advanced microscope technologies for observing the cells. |
Impact | We have observed cells by advanced microscope presenting good results from some of the cells. We also have one collaborative publication. doi.org/10.1016/j.partic.2022.10.014 |
Start Year | 2022 |
Description | Biocrystallisation within designed process |
Organisation | Tianjin University |
Country | China |
Sector | Academic/University |
PI Contribution | We try to understand the protein crystallisation under different conditions, such as shear rate, flow, and soft additives. In my team, we have developed the protein crystallisation technology and training on protein crystallisation technology. |
Collaborator Contribution | We try to understand protein crystallisation under different conditions, such as shear rate, flow, and soft additives. In the collaborative team, they have used different platforms and additives to observe the influence of these parameters on nucleation and protein crystal growth. |
Impact | We have one collaborative publication. doi.org/10.1016/j.ces.2022.117946 |
Start Year | 2021 |
Description | Biomanufacturing with Phage technology |
Organisation | Xi'an Jiaotong University |
Country | China |
Sector | Academic/University |
PI Contribution | Based on the phage technology developed in Xi'an Jiaotong University, we proposed to collaborate on biocrystallisation in biomanufacturing and purification with assistance of phase technology. In our research team, we have tried to cell culture with several phage to investigate the influence of the biocrystallisation process. |
Collaborator Contribution | Based on the phage technology developed in Xi'an Jiaotong University, we proposed to collaborate on biocrystallisation in biomanufacturing and purification with assistance of phase technology. In the collaboration research team, they have tried to design and investigate different phages on different cells. |
Impact | We have a collaborative publication. doi.org/10.3389/fmicb.2021.674415 |
Start Year | 2021 |
Description | Biomanufacturing with Phage technology |
Organisation | Xi'an Jiaotong University |
Country | China |
Sector | Academic/University |
PI Contribution | Based on the phage technology developed in Xi'an Jiaotong University, we proposed to collaborate on biocrystallisation in biomanufacturing and purification with assistance of phase technology. In our research team, we have tried to cell culture with several phage to investigate the influence of the biocrystallisation process. |
Collaborator Contribution | Based on the phage technology developed in Xi'an Jiaotong University, we proposed to collaborate on biocrystallisation in biomanufacturing and purification with assistance of phase technology. In the collaboration research team, they have tried to design and investigate different phages on different cells. |
Impact | We have a collaborative publication. doi.org/10.3389/fmicb.2021.674415 |
Start Year | 2021 |
Description | New technology on Biocrystallisation |
Organisation | Imperial College London |
Country | United Kingdom |
Sector | Academic/University |
PI Contribution | We work together on new technologies protein crystallisation. |
Collaborator Contribution | The collaborator contributes to the training of researchers on some technologies of protein crystallization. |
Impact | We have one collaborative article published. doi.org/10.1021/acs.cgd.9b01447 |
Start Year | 2019 |
Description | Engagement of crystallisation with primary school student |
Form Of Engagement Activity | Participation in an activity, workshop or similar |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Schools |
Results and Impact | We organised an event/activity in a public green field near the primary school after school with about 15 primary school students interested and joining the event in Sep 2022. We presented some crystal samples, such as salt, and sugar, and we give some printed images of protein crystals and small molecular crystals with explanations of the cell cultures. We also showcase that crystallisation in nature. The event was followed by many questions on crystals from primary school students. |
Year(s) Of Engagement Activity | 2022 |
Description | School Open day Lab |
Form Of Engagement Activity | Participation in an open day or visit at my research institution |
Part Of Official Scheme? | No |
Geographic Reach | Local |
Primary Audience | Industry/Business |
Results and Impact | The pharmaceutical industry collaborator joined our school open day and had a lab tour in our biocrystallisation lab. |
Year(s) Of Engagement Activity | 2022 |