Bioreactor Protein Production Primer

This three-day over one weekend course is a comprehensive practical course in bioprocess engineering and the production of recombinant proteins in a fed-batch process. 

A protein production workflow is streamed live, alternating with tutorials, 24 hours a day for the three days.  

Background

Fermentation processes play a very important role in the biotechnology industry. Many proteins are synthesized using microbial fermentation, and a demand for expertise in fermentation processes already exists in the pharmaceutical industry, and is only increasing even more with the rapid development of the alternative proteins food industry worldwide.

Objective

The main objective of the course is to familiarize participants with the challenges of fermentation and of a real-life bioprocess engineering application in the lab sessions using a bioreactor.

The lab work consists of assisting to a real-life protein production session using Escherichia coli carrying a plasmid to recombinantly produce green fluorescent protein (GFP) in a fed-batch process. This can easily be visualized in both the bioreactor and samples by using ultraviolet light.

Audience and Prerequisites

The course is aimed mainly at people who intend to pursue a career in the protein production industry, for instance designing and executing a process to produce alternative proteins for vegan food. It requires undergraduate-level knowledge of biology and chemistry.

Course Organization

The course is organized into six learning units, each composed of a tutorial and a hands-on lab session. During the tutorials and lectures, basic knowledge and relevant parameters are presented to the participants, who then assist to the described experiment as carried out by the instructors in the lab. 

Lab Equipment

The course is performed with a state-of-the-art fermentation platform, including a bench-top bioreactor of standard design with online measurement of relevant reaction parameters such as temperature, pH and pO2 (partial pressure of oxygen).

Digital peristaltic pumps are used to control the flow of feeding solutions in fed-batch mode and the addition of base and acid substances to control the reaction’s pH. All the sensors and the actuators are operated through a computer that is connected to a cloud interface for remote monitoring, including a small camera for visual inspection.

The cloud interface is based on commercially available hardware and custom software we have developed and it is hosted on Google Cloud. It will be streamed live for the whole duration of the course.

Participants assist to every single step of the entire reaction. Starting from the preparation of the seed culture, the setup and installation of the bioreactor, the conduct and control of the reaction, and sampling through the analysis of the online and offline process data, to a final critical evaluation of the reaction and a cost analysis.

Instructors

Julian Ramirez, Biologist, Principal Scientist, Final Foods Inc.

Marco Graziano, Engineer, CEO, Final Foods Inc.

Syllabus and Schedule

The course is in three days and it is divided into six learning units. An overview of the schedule is given in the diagram below:

Learning Units

Unit 1:

Lecture: Fundamentals (Principles and tasks)

Getting to know the bioreactor

Collecting basic information for the production of recombinant proteins. Preparing instruments and discussing safety measures.

Unit 2:

Tutorial: Reaction setup (and preparations)

Setting up the bioreactor

Creation of a work plan, sensors calibration.

Unit 3:

Tutorial: Process and workflow (and process control)

Operating the bioreactor part 1

Estimate process parameters 

Unit 4:

Tutorial: Process control

Operating the bioreactor part 2

Monitoring the process and compare with estimates

Unit 5:

Tutorial: Analysis and calculations

Results preparation and measurements

Estimate reaction outcome

Unit 6:

Tutorial: Scientific documentation and writing

Documentation of the reaction and results preparation

The first day the course starts in the evening with a lecture which gives an introduction to the topic and outlines principles and tasks for the participants for the following days. During the lecture, basic knowledge and relevant parameters are collected by the instructors, which are required to perform the experiment. 

Lab work is performed by the instructors and streamed live 24 hours a day.  Each of the following 2 days starts with a tutorial to discuss and brief the participants on what specific steps are necessary on each individual day.