Bacterial Recombinant Protein Expression
As a company with rich experience in developing custom antibodies to difficult targets, Maine Biotechnology Services brings the specialized perspective of antibody development professionals to recombinant protein development services. When you choose MBS to develop recombinant antigen for your hybridoma development, the end use application of your antibodies are considered during every phase of antigen production. When protein production is complete, MBS provides a seamless and time-saving transition to immunizations quickening the ultimate objective of obtaining antibodies that meet your requirements.
- Bacterial Recombinant Protein Production Initiation Checklist: To successfully develop a recombinant protein that meets the customer’s needs, communication is key. This checklist is the customer’s first opportunity to provide MBS with details necessary to strategize an optimal approach.
- Technical Meeting: Projects begin with a conference call with the Recombinant Protein Development team to clarify the goals and customize the project for success. If the end use of the protein is as antigen for antibody development, the meeting can include members of the Hybridoma Development team. This provides the opportunity for collaborative antigen to monoclonal antibody strategizing at the earliest possible stages.
- Pre-Project Protein Analysis: Individual protein characteristics and sequence are analyzed in order to assess the suitability for bacterial expression and anticipate potential risks to guide protein production strategy.
Expression Strategy and Obtaining Expression Construct
- Devise Expression Strategy: Based on the analysis of the protein and goals of the project.
- Obtain Expression Construct: Either provided by the customer or MBS will facilitate the gene synthesis and cloning into expression vector of choice. His, GST, and MBP tags standard.
- Sequence Verification: After storage glycerol stocks have been generated, the protein’s DNA sequence will be verified prior to expression.
Small Scale Expression & Purification Evaluation
- Expression constructs are transformed into multiple host strains and a small scale expression, isolation, and purification performed to determine optimal parameters for large scale production.
- If expression levels are low or the protein is insoluble, the customer will be contacted promptly to discuss recommendations.
- Bacterial recombinant protein expression carries certain risks such as low expression levels or insolubility. To address this, MBS uses a parallel approach to evaluate multiple expression conditions and move the project forward as quickly as possible.
Estimated time = 2 weeks
Large Scale Production & Purification
- One liter expression based on conditions identified in Phase I. Purification performed using automated purifier (AKTA).
- Purity is determined via an Experion automated chip-based electrophoresis.
- Protein and reports are delivered directly to the customer or transferred to the Hybridoma Development group to be used in an antibody development project.