Industrial chromatography

Relatedness (affinity) chromatography is based on a reversible interaction between a protein and a specific ligand coupled to a solid carrier. This type of chromatography is a very efficient biomolecule separation technique. This is metal chelating chromatography designed separation of histidine-tagged proteins from compounds (protein purification matrix). The separation is triggered by using affinity ligands on the sorbent matrix. The ligands can be a substrate or coenzyme. Only proteins are binding to the ligands. This interaction is of a specific nature. All other proteins go to the eluate. Relatedness (affinity) chromatography is characterised by high specificity and allows to achieve protein purification levels in the order of several thousand-fold. Protein A ligand sorbents for purification of monoclonal antibodies.

Proteins are complex molecules that can have positively or negatively charged surface areas. The most widely used method to produce a pure protein is ion exchange (IEX) chromatography. It separates molecules based on the charge distribution on their surface with functional groups of the chromatography media carrying the opposite charge. Ion exchange chromatography is based on the exchange reaction between the buffer ions and mobile sorbent ions, occurring when the buffer flows through the cationic or anionic column. By choosing the right chromatographic conditions (e.g. salt concentration, buffer system and pH), proteins can be separated from impurities. In common practice, bound molecules are selectively eluted by step-wise or continuously increasing concentration of a monovalent salt. Impurities can be bound to the resin or membrane in a flow through mode. Merck sorbents for ion exchange chromatography: Fractogel®, Eshmuno®.

The advantage of reverse phase chromatography is that it delivers high levels of purity in a rapid and economical manner. Preparative column chromatography plays an important role in purifying valuable compounds in research or production. If silica gel for advanced normal-phase and reversed phase chromatography is required, we recommend using the irregular shaped LiChroprep® or the perfect spherical PharmPrep™ - highly versatile materials providing fast, effective and reproducible separation.

For reverse phase chromatography the quality of sorbent is of tremendous importance. One of the most important factors in chromatography is selectivity. It is decisive in determining the quality and the reproducibility of a chromatographic separation. Merck's inorganic sorbents will deliver reproducible results within narrow band ranges. This provides the highest degree of reliability for chromatographic separations and purifications in the laboratory, pilot plant and production.

Protein purification and final cleaning depends on chosen chromatography techniques that isolate targeted proteins and remove all remaining impurities. This method is based on differing hydrophobic interaction of particles of different molecular mass between the stationary and mobile phase of the chromatography column.

Hydrophobic interaction chromatography is a perfect choice for protein purification. This technique is based on a hydrophobic interaction between hydrophobic ligands and hydrophobic groups of the protein molecules. High-concentration salt buffer helps to separate the needed protein in a more efficient manner. 

Molecular sieve chromatography (gel filtration) is designed for final protein purification. The requirements to the buffer are quite simple. Native or recombinant proteins, viruses and plasma of biological origin can be easily purified with molecular sieve chromatography columns.

Choosing the right pre-packed chromatography columns is crucial for successful separation and purification. The next important step is packing your sorbent into the column (packing control and validation). Merck offers a modern-day solution, Chromabolt® pre-packed chromatography columns.  Available in a standard or tailor-made format. Designed for use in a laboratory, low-scale pilot or continuous production. With Chromabolt® pre-packed columns, you can spend your time on manufacturing your products instead of on laboriously packing chromatography resins. Available in different sizes and sorbent combinations.

It is paramount to ensure stability and cleanness in each step of every production process. That is why we offer a wide range of chemical substances, from salts to concentrated solutions. All preparations are ready-to-use and packed in single-use packs or rigid plastic containers. Merck EMPROVE® bio and EMPROVE® exp products are provided with an extensive quality assurance documentation. EMPROVE® products will make help you to work faster and more cost-efficient.

Buffer is used in nearly each stage of any biopharmaceutical manufacturing process, which is why buffer quality impacts the success of the process. Merck offers high-quality buffer substances, compatible with all commonly used buffer systems, such as phosphates, acetates, and organic buffers TRIS, HEPES, MES.

Production processes are developed to obtain reduced microbiological and endotoxin contamination. Moreover, enzyme activity (DNase, RNase, protease) of most buffers is still unknown, but crucial for biochromatography. To this effect qualification kits with samples from three batches can be used.

Vast amounts of cleaning-in-place (CIP) substances are used in the biopharmaceutical manufacturing. Some manufacturers order on-demand made cleaning substances. Merck has developed high-quality solutions for cleaning and storing of chromatography media. These are Fractogel® and ProSep® sorbents and Pellicon® cassettes. All products fulfil the highest requirements to purity and reliability, and effectively prevent contamination of your valuable purification equipment. All solutions are manufactured under GMP conditions.