Affinity Purification

Pre-Activated Monoliths for Immobilization of Affinity Chromatography Ligands

BIA Separations manufactures a full line of pre-activated monoliths media and services to support your affinity chromatography needs. These products enable you to covalently immobilize peptides, proteins, oligonucleotides, and carbohydrates. Or if you would prefer, we can immobilize them for you on a custom basis and scale them up as your needs require.

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As with non-affinity applications, monoliths offer disproportionately higher capacity for affinity purification of large bioproducts; 10–100 times higher capacity than porous particle columns. Those large bioproducts can be large proteins like IgM. They can be oligonucleotides, including both RNA and DNA. Or they can be virus particles, cellular organelles, or vesicles like exosomes.

Monoliths also enable synthesis of superior enzyme bioreactors because their is no diffusive lag for the time it takes for the substrate to diffuse into and out of pores. Unrestricted enzyme-substrate contact is enabled by the large convective channels and convective mass transport from them. Reaction kinetics very closely approach and sometimes exceed the kinetics observed in free solution.

BIA Separations pre-activated affinity products come in a variety of sizes and formats. CIMmic™ monoliths with 100-200 µL beds are ideal for initial screening. This includes situations where you might need to make capacity estimates as part of characterizing various ligands and the amount of source material is limited. The ability of monoliths to support flow rates 20–50 times higher than porous particles further favors rapid evaluation.

CIMac™ affinity monoliths with 100 µL bed volumes are ideal for analytical scale applications. The CIMmultus™ line is represented by radial flow monoliths as small as 1 mL for method development, presently up to 8 L industrial scale units, and soon up to 40 L. BIA Separations also offers CIM™ 96-well plates for high-throughput robotic sampling.

Table 1: BIA Separations pre-activation chemistries and their target residues
ALD, aldehyde

Binds bioproducts by their primary amino residues

CDI, carbonyl diimidazole

Binds bioproducts by their primary amino residues

EDA-GLU, glutaraledehydeLong spacer with binding to primary amino residues
HDZ, hydrazide

Binds bioproducts by their carbohydrate residues, enables site-directed immobilization of IgG

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Site-directed imobilization of IgG by hydrazide chemistry.

Most often used