Creative Biostructure

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MagHelix™ Protein Purification

Creative Biostructure has developed an advanced purification platform to provide protein samples with "crystallographic grade" purity (i.e., >95%) and ready for crystallization and other high quality assays. With this platform, we offer services to meet your specific needs in an efficient and economical way. Our continuous fermentors and large-size chromatography systems allow production of high purity proteins ranging from milligram to gram scales.

Types of proteins:

  • Native proteins from natural sources
  • Tagged recombinant proteins
  • Antibodies
  • Enzymes

Expression Systems:

  • Bacterial
  • Baculovirus
  • Yeast
  • Mammalian Cell

Protein Purification

With our expertise in protein chemistry, analytical chemistry and functional assays, Creative Biostructure is able to deliver efficient and cost-effective solutions in large-scale protein purification and process development. We also undertake the development of challenging proteins, such as co-expression or assembly of higher order complexes. Depending on the project requirements and the nature of the proteins, purification protocols can be either de-novo designed by us or obtained from customers or literature. In Creative Biostructure, proactive efforts have been put into literature reviews of the latest protocols available. Our protein purification methods include affinity chromatography with various tags, protein-specific affinity chromatography, etc. And the final goal is to accomplish a homogeneous and biologically active protein preparation that is critical for crystallization, protein-drug assays and other related studies.

Protocols:

  • De-novo protocols
  • Existing protocols from customers or published sources
  • Optimized protocols for various expression systems (e.g., bacterial, yeast, etc.)

Purification Systems:

Affinity chromatography is a separation method based on a specific binding interaction between an immobilized ligand and its binding partner. Examples include antibody/antigen, enzyme/substrate, and enzyme/inhibitor interactions.

Ion Exchange Chromatography relies on charge-charge interactions between the proteins in your sample and the charges immobilized on the resin of your choice, including Anion Exchange Chromatography (AEC) and Cation Exchange Chromatography (CEC).

Size exclusion chromatography is one of the HPLC separation modes. When dissolved molecules of various sizes flow into the column, smaller dissolved molecules flow more slowly through the column because they penetrate deep into the pores, whereas large dissolved molecules flow quickly through the column because they do not enter the pores. Consequently, larger molecules elute from the column sooner and smaller molecules later, which effectively sorts the molecules by size.

HIC is particularly suitable for samples precipitated with ammonium sulfate or eluted in high salt concentrations since high ionic strength buffers enhance the hydrophobic interaction. The interaction is enhanced by buffers with high ionic strength, which makes HIC an excellent purification step after ammonium sulfate precipitation or after elution in high salt during ion exchange chromatography (IEX). HIC is well-suited for the capture or intermediate steps in a purification scheme.

RPC exploits reversible adsorption of biomolecules according to their hydrophobicity under conditions where the stationary phase is more hydrophobic than the mobile phase. RPC is mainly used for high resolution separation in the analysis of proteins, peptides, and nucleic acids. The technique is highly recommended for applications such as peptide mapping or purity checking and is often used for final polishing of oligonucleotides and peptides.

Tangential flow filtration (TFF) is a rapid and efficient method for separation and purification of biomolecules. It can be applied to a wide range of biological fields such as immunology, protein chemistry, molecular biology, biochemistry, and microbiology. TFF can be used to concentrate and desalt sample solutions ranging in volume from 10 mL to thousands of liters. It can be used to fractionate large from small biomolecules, harvest cell suspensions, and clarify fermentation broths and cell lysates.


Protein Analysis:

  • Protein Assays (e.g. A280, BCA, etc.)
  • Native-PAGE/SDS-PAGE (Coomasie-stained, Silver-stained)
  • Western Blot/Dot Blot
  • ELISA