How does perfusion chromatography work?

Answer 1

Perfusion chromatography is a technique

arised

to overcome the problem associated with mass transfer in the separation

of large

molecules such as proteins by high-performance

liquid chromatography (HPLC). Perfusion media are constituted by

two set of pores:

•

throughpores

(6000--8000 A

°

)

•

diffusive pores

(800--1500 A

°

) which enable better access

of.

Macromolecules

to the inner of the particle by the combination of convective and diffusive flow.

General

characteristics of

perfusion

chromatography

In conventional liquid chromatography (diffusion chromatography) molecules move to the outer surface of the stationary phase particles by convection which constitutes a rapid step Nevertheless, transport of molecules through a stationary phase particle occurs by molecular diffusion which constitutes a very slow process, especial

ly

in the case of peptides and proteins that present high molecular weights and low diffusion coefficients . As a consequence, a significant increase in bandspreading (loss in resolution) could occur.

In perfusion chromatography (flow-through particle chromatography), stationary phase particles

used are designed to enable a better access of molecules to the inner of these through two classes of pores:

throughpores

(6000–8000 A

°

), which cross

damthe

stationary phase particle from side to side and allow the transport of molecules into the interior of the particle by convective flow and

diffusive pores

(800–1500 A

°

), interconnecting the

throughpore

network and enabling the transport by diffusion In this way, molecules travel by convection

through the column to the stationary phase particle, such as in conventional chromatography. Since there, molecules cross the stationary phase particles by means of a combination of convective and diffusive transport, thus, accelerating the transport of molecules through the particle. This singular bimodal structure significantly enhances mass transport of molecules...