A GLOBAL COLLABORATION FOR EQUITABLE ACCESS TO BIOTECHNOLOGY
Local Manufacturing of Reagents
Wed 10 June, 14:00-17:00 GMT
Virtual symposium featuring talks from projects manufacturing enzymes and reagents in Africa, Asia, Latin America and beyond PLUS breakout groups on aspects of scaling up, regulation, intellectual property, supply chains and more. Participate to share your experiences and join Reclone Network working groups
LOCAL MANUFACTURING OF REAGENTS SCHEDULE
|14:00||Introduction to the Reclone Network|
|14:05||The Research in Diagnostics DNA Collection: an open source toolkit for production of enzymes for diagnostic research||Chiara Gandini, University of Cambridge, UK|
|14:30||Lightning Talks Session 1|
|One-step RT-PCR from locally produced M-MLV + pfu-Sso7d||Anibal Arce, Pontificia Universidad Católica de Chile, Chile|
|Optimization of a low-cost RNA extraction protocol for SARS-Cov2 based on magnetic beads||Alejandro Caro, AGROSAVIA, Colombia|
|ViroDet: a community effort on COVID-19 diagnostics for Mexico and Latin America||Ricardo Chavez, Scintia, Mexico|
|14:50||Breakout Sessions 1|
|1a - What do you want to know about enzyme expression? Help compile an FAQ for the Research in Diagnostics Collection|
|1b - ISO 101: How do new reagent manufacturers navigate quality assurance, diagnostic regulation and certification?|
|1c - Reimagining the reagent supply chain: your top ideas to reach equitable global distribution|
|1d - Financing and Partnerships for local reagent production|
|15:20||Scaling up local manufacturing of reagents||Cassi Henderson, University of Cambridge, UK|
|15:35||Lightning Talks Session 2|
|Advances by ReClone_Argentina||Alejandro Nadra, University of Buenos Aires, Argentina|
|BEARMix - open-source master mix for 1-step RT-qPCR||Thomas Graham, UC Berkeley, USA|
|Quality Management Systems for enzyme production in Cameroon||Nadine Mowoh, MboaLab, Cameroon|
|15:55||Biotechnology and the Public Domain: how to find out if technology is free and open||Linda Kahl, SciScript & Beneficial Bio|
|16:05||Breakout Sessions 2|
|2a - Learning together with RT-LAMP: how do we build collaboratively and openly in a crowded space?|
|2b - Making IP work for equity: how can we work with lawyers to make freedoms more visible|
|2c - Getting reagents around the world: framework for a community approach to mapping the legal and biosafety requirements.|
|2d - Low-cost enzyme purification: what have you tried, what has worked?|
|16:35||Reports on breakout sessions|
|16:55||Summary and next steps to establish Reclone working groups|
|17:00||Meeting closes - online room will be open for continued discussion|
Please apply to present a 5 min lightning talk on your work or suggest a breakout group topic using the registration form below! The meeting link will be emailed to registered participants in advance.
The Research in Diagnostics DNA Collection: an open source toolkit for production of enzymes for diagnostic research
Chiara Gandini, University of Cambridge, UK
Abstract: Enzymes are the heart of each and every diagnostic test. The Research in Diagnostic DNA Collection wants to provide researchers worldwide with an open source toolkit to produce their own enzymes and streamline their research in diagnostics. The collection includes 1) ready-to-use expression cassettes to produce essential enzymes among which RT, Bst-LF, RPA enzymes, RNA inhibitors and Sars-CoV-2 RNA positive and negative controls, 2) DNA modules to build up expression cassettes tailored to your lab’s needs with a choice between 20 genes, 6 promoters, 10 purification tags and 6 cleavage sites, among others 3) an extensive but not exhaustive repository of laboratory protocols as a guide on how to express some of the enzymes of the collection. We will present the collection and its resources, and explain how to use it. We will be keen to hear your thoughts and suggestions on how we can make it more used and useful. This project is a collaboration between the Open Bioeconomy Lab, Fernan Federici’s lab, OSNVancouver and FreeGenes.
Mapping scale-up pathways for local manufacture of affordable diagnostics
Cassi Henderson, University of Cambridge, UK
Abstract: To make the transition from ‘bench to bedside’ for locally manufactured diagnostics, scale-up will be essential. Scale-up is a multidimensional process that encompasses not only technology and manufacturing developments but also business and value chain creation. This talk will cover some established frameworks for managing the dimensions of scale-up and explore specific influencing factors and impact areas relevant to the local production of diagnostics.
Biotechnology and the Public Domain: how to find out if technology is free and open
Linda Kahl, SciScript & Beneficial Bio
Abstract: The patent system is often associated with exclusivity and monopoly, but in fact is one of the best mechanisms for building the public domain. Examples of biotechnologies that have entered the public domain via the patent system include the recombinant DNA technology developed by Herbert Cohen and Stanley Boyer, the polymerase chain reaction developed by Kary Mullis, and the use of green fluorescent protein for monitoring gene expression developed by Martin Chalfie.
The quid pro quo of the patent system is to provide a limited period of exclusivity in exchange for disclosure of the invention to the public. Once the period of exclusivity has ended, the invention enters the public domain where it is available for anyone to use without charge. In this session you will learn how to determine whether or not a patent has expired, or been abandoned, and has entered the public domain.
One-step RT-PCR from locally produced M-MLV + pfu-Sso7d
Anibal Arce, Pontificia Universidad Católica de Chile, Chile
Abstract: As many other countries, Chile faces a significant challenge in the availability and high costs of proprietary enzymes for RT-qPCR diagnostics of SARS-CoV-2. Here, we present our attempts to locally produce an enzyme kit for this purpose.
We first standardized common protein over-expression and purification conditions for DNA polymerases (Pfu-Sso7d, RTX) and reverse transcriptases (M-MLV, Mashup) that minimize unnecessary reagent usage and avoid the risk of protein aggregation while maximizing protein yields, obtaining 8 mg of Pfu and 4 mg of M-MLV per L of culture.
Due to the lack of plasmids for in vitro transcription of positive RNA controls, we designed 5 different plasmids for genetic parts of SARS-CoV-2 that are useful for most published diagnostic tools to date. We also included two plasmids for the controls for N proteins of SARS1 and MERS. Those plasmids have been included in the open Diagnostic collection.
Then, we standardized a protocol for the polymerase activity of Pfu and M-MLV reverse transcriptase activity independently. Later, to simplify protocol in order to use both enzymes in a one-step reaction to optimize the use of these enzymes in SARS-CoV-2 detection. We developed a one-step protocol to do RT-qPCR where both enzymes together are used to directly amplify RNA from in vitro transcription. We were able to successfully amplify through RT-qPCR the N gene from Sars-CoV-2 using 3 different pairs of primers. Future work considers the assessment of the limit of detection of this method and its optimization to reach diagnostics standards and the development of in-house buffers for these reactions.
Contact: Fernan Federici, firstname.lastname@example.org/ email@example.com