Protein Construct Design
Explore the first step with the cutting-edge world of Protein Construct Design at Eurofins CALIXAR. Our expert services, utilizing FBDD and SBDD, deliver fully native, active custom proteins with a focus on stability and functionality. From membrane proteins to soluble targets, our high-quality process ensures maintained structure and integrity. Discover the power of proprietary proteins and elevate your research with precision. Partner with us for excellence in drug discovery – your key to unmutated, untruncated targets awaits.
Foundation of success: strategic protein construct design
Protein construct design marks the crucial initiation of protein production, laying the groundwork for successful research or therapeutic endeavors. The process begins by clearly defining the objective of the protein, whether it's intended for structural elucidation, functional assays, or drug development. The selection of the target protein follows, with careful consideration given to its biological function, structure, and relevance to the study's goals. Subsequently, structural elements are meticulously assessed, identifying key domains or regions essential for the protein's function. This strategic planning ensures that the amino acid sequence is optimized for enhanced expression, solubility, and stability, setting the stage for a streamlined and effective protein production process.
Protein construct design process: precision from concept to completion
The process of gene design in protein production involves several key techniques, each playing a crucial role in optimizing the expression, functionality, and yield of the target protein.
1. Gene synthesis: building the blueprint
The process begins with gene synthesis, where the DNA sequence encoding the target protein is artificially created. This step allows for the optimization of codon usage, removal of unwanted elements, and incorporation of specific features such as affinity tags or regulatory elements. Eurofins CALIXAR employs advanced gene synthesis techniques to ensure the generation of highly customized and optimized gene sequences.
2. Codon optimization: maximizing expression efficiency
Codon optimization involves the strategic alteration of codons in the gene sequence to match the preferred codon usage of the host expression system. This optimization enhances translation efficiency and can significantly improve protein expression levels. Eurofins CALIXAR utilizes cutting-edge tools to fine-tune codon usage for optimal protein production.
3. Vector design: creating the delivery system
The synthesized gene is then inserted into a vector, which serves as the delivery system for introducing the gene into the host organism for expression. Vector design involves choosing the appropriate expression vector, promoter, and other regulatory elements to achieve the desired level and timing of protein expression. Eurofins CALIXAR customizes vector designs based on the specific requirements of the target protein and the expression host.
4. Promoter selection: controlling expression levels
Promoters play a critical role in controlling the rate of transcription and, consequently, the expression level of the target protein. Eurofins CALIXAR carefully selects promoters that align with the desired expression profile, taking into consideration factors such as inducibility and strength.
5. Inclusion of affinity tags: simplifying purification
Affinity tags, such as His-tags or GST-tags, are often incorporated into the gene design to facilitate easy purification of the expressed protein. These tags enable efficient isolation of the target protein from the complex cellular milieu. Eurofins CALIXAR expertly integrates affinity tags into the gene design, ensuring minimal interference with the protein's structure and function.
6. Signal peptide addition: guiding protein secretion
For proteins intended for secretion, the addition of a signal peptide is crucial. This sequence directs the protein to the secretory pathway, allowing for extracellular expression. Eurofins CALIXAR considers the nature of the target protein and the desired cellular localization when incorporating signal peptides into gene designs.
7. Codon harmonization: enhancing translation efficiency
Codon harmonization involves aligning the codon usage of the target gene with the tRNA pool of the host organism. This further enhances translation efficiency and reduces the likelihood of translational bottlenecks. Eurofins CALIXAR employs sophisticated strategies for codon harmonization to maximize protein production.
Highlight
Protein tagging excellence - Advanced techniques for precise protein tagging, enhancing functionality and traceability for various applications.
Lead sequences for optimal expression - Collaborative identification and design of lead sequences, forming a solid foundation for highly optimized protein expression.
Precision in protein release - Clever incorporation of HRB3C, TEV, and Thrombin cleavage sequences, facilitating seamless and precise release during downstream processes.
Preservation of unmutated and untruncated targets - Emphasis on designing constructs that maintain the target protein in its unaltered and complete form.
Affinity tags
Precision in protein purification with affinity tags—molecular handles that streamline the process by offering specific binding sites for purification reagents. This precision guarantees the isolation of only the desired protein, eliminating contaminants. Tailored to diverse needs, our affinity tags ensure optimal performance, whether your project requires gentle elution conditions or robust purification protocols.
Commonly used affinity tags in recombinant protein production:
6xHistidine Tag (His-tag) Widely recognized for its simplicity and efficiency, the His-tag binds to metal ions for purification via immobilized metal affinity chromatography (IMAC).
Maltose-Binding Protein Tag (MBP): Ideal for proteins prone to aggregation, MBP facilitates gentle purification via amylose or starch affinity chromatography.
Glutathione S-Transferase Tag (GST): Harness the power of GST for efficient purification through glutathione affinity chromatography, suitable for a wide range of protein targets.
Strep-Tag: Explore the versatility of Strep-Tag, binding with high affinity to Strep-Tactin for efficient purification.
FLAG Tag: For those who require an easily detectable tag, the FLAG tag is recognized by anti-FLAG antibodies, streamlining both purification and detection.
Proprietary proteins for exclusive insights
Eurofins CALIXAR takes pride in offering proprietary proteins, giving you exclusive access to resources that set your research apart. Benefit from our expertise and gain unique insights that drive your projects forward.
Partner with Eurofins CALIXAR for Protein Construct Design services that go beyond expectations. Elevate your research with fully native, active custom proteins designed to meet the highest standards of quality and precision.
Contact us today to discuss how our Protein Construct Design services can amplify the impact of your research.
Secure and boost
your discovery programs
Starting from native material or recombinant systems, we succeed with all types of proteins: Kinases, Phosphatases, Ubiquitins, Epigenetic Proteins, GPCRs, Ion Channels, Transporters, Receptors and Viral Proteins.
