Protein Expression Services
Aviva provides customized solutions to each protein expression project. Our expert scientific staff has experience with difficult proteins and have produced more than 2,500 recombinant proteins. Whether your needs are small scale research or large scale cGMP, we can apply our experience to meet your needs. Services include E. coli expression, Mammalian expression, Yeast expression, and Antigen Preparation. Please use our convenient online Protein Expression Quote Form if you would get a quote or schedule a call to discuss your needs.
| Contact us and receive a quote within 24 hours. | Email: info@avivasysbio.com Phone: 858-552-6979 |
E. coli Protein Expression Services
Aviva recombinant protein expression in E. coli includes antigen production, ligand preparation, and proteins for binding assays, cell based assays, enzymatic assays, and assays for vaccine and therapeutic development. Our proprietary platforms can produce functional recombinant proteins, domain specific fragments, fusion proteins, and protein complexes. We understand the need for active proteins, free of impurities, to produce clean reproducible results.
View our ISO13485 compliant protein development, validation, and production options.
The following is our E. coli custom protein expression development template:
| Order | Process | Details |
|---|---|---|
| Step 1 | Bioinformatic analysis, domain selection and vector design | Aviva conducts bioinformatic analysis on target(s) and recommends an expression strategy to each client. Sequence verified cDNAs are provided by the client, supplied by Aviva, or are purchased from available commercial sources. |
| Step 2 | Prepare subcloning plasmid and construct expression vectors | Using Aviva's vector system with the most appropriate strategy to construct vectors. |
| Step 3 | Target sequence analysis and confirmation | Conduct DNA sequencing and analysis to confirm correct coding and flanking sequence. |
| Step 4 | Cell transformation and expression clone preparation | Use Aviva's proprietary expression cell lines and prepare expression cell clone. |
| Step 5 | Protein production and purification | Express protein in a small scale culture, collect and lyse cells, isolate and purify protein. If the protein is not soluble, then the processes of solubilization and refolding will be initiated. |
| Step 6 | Quality control test | Conduct SDS-PAGE analysis and generate QC report. |
| Step 7 | Scale-up production and quality control test | Produce and purify target proteins at a large scale. |
| Step 8 | Characterization of target protein (including functional studies) | 1. Determine purity in conjunction with client specifications. 2. Employ a functional assay method provided by client or to be specifically developed. 3. Utilize immunoassays such as western blot analysis, ELISA, etc, after antibodies are produced or provided. |
Each protein has its unique biochemical feature in the expression process. Aviva's team will work with the client to develop the best strategy and use the most cost-effective process.
Purification Methods
- Affinity Chromatography
- Anti-Flag
- Glutathione Sepharose
- Heparin Sepharose
- Ni-NTA
- Protein A
- Protein G
- Streptavidin Sepharose
- Talon resin
- Ion Exchange Chromatography
- ANX
- CM Sepharose
- DEAE Sepharose
- Mono-Q
- Q Sepharose Fast Flow
- Q Sepharose XL
- Resource Q
- SP Sepharose
- Hydrophobic Interaction Chromatography
- (CH2)n Sepharose
- Phenyl Sepharose
- Size Exclusion Chromatography
Optional Services
- Tag Removal
- Endotoxin Removal
- Protein solubilization and refolding
Mammalian Protein Expression Services
Aviva mammalian expression systems generate high-yields of mammalian proteins. These expression systems are often suggested for proteins requiring mammalian post-translational modifications. Aviva has a great amount of experience producing monoclonal antibodies or recombinant proteins from a wide range of cell types including CHO, hybridomas, various mouse myeloma, and human cell lines. Aviva performs a comprehensive analysis of your project in order to identify the best expression platforms with appropriate scalability, yield, and functionality. Our service includes bioinformatics analysis, domain selection, vector design, tag selection, sequence confirmation, cell transfection, cell amplification, purification by tag selection, yield optimization, scale up processing, and protein characterization.
View our ISO13485 compliant protein development, validation, and production options.
The following is our mammalian protein expression development template:
| Order | Process | Details |
|---|---|---|
| Step 1 | Bioinformatic analysis, domain selection and vector design | Aviva conducts bioinformatic analysis on target(s) and recommends an expression strategy to each client. Sequence verified cDNAs are provided by the client, supplied by Aviva, or are purchased from available commercial sources. |
| Step 2 | Construction of expression vectors | The target sequence will be cloned into Aviva's vector system. Based on the specific target protein, the sequence can be inserted into an expression cassette for a secreted or non-secreted protein. Aviva will conduct DNA sequencing and analysis to confirm correct coding and flanking sequences. |
| Step 3 | Cell transfection, selection, and cloning of target-expressing cells | The protein can be produced by transient burst or stable cell line expression depending on the client's specific needs. The expression of the target protein will be confirmed by SDS page or Western blot using an anti-tag antibody or client-provided antibody against the target protein. |
| Step 4 | Protein production and purification | Aviva will express the protein in a scale of up to 4x T-75 culture flasks or equivalent. The protein will be purified using a tag-assisted affinity column or other chromatographic methods. Protein samples may be provided to clients if the protein function needs to be confirmed. |
| Step 5 | Scale-up production and quality control test | Aviva will produce the target protein(s) at large scale. The scale of production depends on the level of protein expression, the yield of purification, and the requirements of the client. |
| Step 6 | Characterization of target protein (including functional studies) | 1. Determine purity in conjunction with client specifications. 2. Employ a functional assay method provided by client or to be specifically developed. 3. Utilize immunoassays such as western blot analysis, ELISA, etc, after antibodies are produced or provided. |
Each protein has its unique biochemical feature in the expression process. Aviva's team will work with the client to develop the best strategy and use the most cost-effective process.
Purification Methods
- Affinity Chromatography
- Anti-Flag
- Glutathione Sepharose
- Heparin Sepharose
- Ni-NTA
- Protein A
- Protein G
- Streptavidin Sepharose
- Talon resin
- Ion Exchange Chromatography
- ANX
- CM Sepharose
- DEAE Sepharose
- Mono-Q
- Q Sepharose Fast Flow
- Q Sepharose XL
- Resource Q
- SP Sepharose
- Hydrophobic Interaction Chromatography
- (CH2)n Sepharose
- Phenyl Sepharose
- Size Exclusion Chromatography
Optional Services
- Tag Removal
- Endotoxin Removal
- Protein solubilization and refolding
Yeast Protein Expression Services
Aviva's yeast expression platform provides a flexible, controllable, and effective solution for protein expression needs. Aviva's various yeast expression platforms have been well characterized and are validated to perform many post-translational modifications. Aviva offers an initial assessment of your project and makes recommendations regarding expression platforms with appropriate scalability, yield, and functionality. Our approach includes bioinformatics analysis, domain selection, vector design, tag selection, sequence confirmation, cell transfection, cell amplification, purification by tag selection, yield optimization, scale up processing, and protein characterization.
View our ISO13485 compliant protein development, validation, and production options.
The following is our yeast protein expression development template:
| Order | Process | Details |
|---|---|---|
| Step 1 | Bioinformatic analysis, domain selection and vector design | Aviva conducts bioinformatic analysis on target(s) and recommends an expression strategy to each client. Sequence verified cDNAs are provided by the client, supplied by Aviva, or are purchased from available commercial sources. |
| Step 2 | Prepare subcloning plasmid and construct expression vectors | Project design includes Aviva's vector system with the most appropriate strategy to construct vectors. If certain specific vectors are requirgb(170,0,0), then custom vectors will be included in the project design. |
| Step 3 | Target sequence analysis and confirmation | Conduct DNA sequencing and analysis to confirm correct coding and flanking sequence. |
| Step 4 | Cell transformation and expression clone preparation | Aviva will utilize proprietary expression cell lines and prepare expression cell clone. If specific cell lines (e.g. mutants) are needed for accommodating the need for recombinant protein expression and selection, Aviva's team will discuss with client to make specific arrangements for the project. |
| Step 5 | Protein production and purification | Express protein in a small scale culture. Fermentation broth or cell lysate (prepargb(170,0,0) by using microfluidizer) will be collected for filtration and concentration. Expressed protein will be further isolated and purified according the protocol provided by client or developed by Aviva. If the protein is not soluble, processes of solubilization and refolding will be initiated. |
| Step 6 | Quality control test | Conduct SDS-PAGE analysis and generate QC report. |
| Step 7 | Scale-up production and quality control test | Produce and QC target proteins at a large scale according to client requirements. |
| Step 8 | Characterization of target protein (including functional studies) | 1. Determine purity in conjunction with client specifications. 2. Employ a functional assay method provided by client or to be specifically developed. 3. Utilize immunoassays such as western blot analysis, ELISA, etc, after antibodies are produced or provided. |
Each protein has its unique biochemical feature in the expression process. Aviva's team will work with the client to develop the best strategy and use the most cost-effective process.
Purification Methods
- Affinity Chromatography
- Anti-Flag
- Glutathione Sepharose
- Heparin Sepharose
- Ni-NTA
- Protein A
- Protein G
- Streptavidin Sepharose
- Talon resin
- Ion Exchange Chromatography
- ANX
- CM Sepharose
- DEAE Sepharose
- Mono-Q
- Q Sepharose Fast Flow
- Q Sepharose XL
- Resource Q
- SP Sepharose
- Hydrophobic Interaction Chromatography
- (CH2)n Sepharose
- Phenyl Sepharose
- Size Exclusion Chromatography
Optional Services
- Tag Removal
- Endotoxin Removal
- Protein solubilization and refolding
Antigen Preparation Services
One of the determining factors in quality antibody development is quality antigen preparation. We focus on preparation of high-quality antigen for ensuring high-quality antibody development.
- Using our bioinformatics core we determine the best immunogenic domain for antibody generation.
- Using our proprietary fusion domain and vectors we express and purify recombinant antigens in the most cost-effective manner.
- Aviva can provide full services, with the customer providing a target protein accession number and Aviva delivering high-quality polyclonal or monoclonal antibodies.
There are different natures and great variations of antigens in type, form, size, availability, etc. Each type of antigen has its own utility, advantages and disadvantages.
The following details and compares various considerations for antigen development:
| Comparison of Antigens | ||||
|---|---|---|---|---|
| Consideration | Native Proteins | Recombinant Proteins | Peptides | Other |
| Size and form | Full-Length Vary in size | 50-300 aa (Domain) or Full-Length | 6-20 aa | Small molecule, macromolecule whole cell or microorganism |
| Conjugation (Carrier) | Not Needed | Not Needed | Needed | Needed for small and macromolecule |
| Fusion Tag | No | Optional | No | Optional |
| Immunogenicity | Best | High | Fair | Fair |
| Success Rate for Antibody Generation | 80-95% | 60-80% | 30-60% | 40-60% |
| Suitable as Ligand | Best | Good | Fair | Vary |
| Immuno-reactivity | Good | Good | Vary | Vary |
| Reproducibility | Best | Good | Good | Vary |
| Cost | High | Middle | Low | Vary |
Aviva's antigen technology specializes in the three major types:
Purification Methods
- Native Proteins
- Peptides and Conjugates
- Recombinant Proteins