Cell microfiltration

For industrial biotransformations, catalyst recovery and reuse are major issues. This may be desirable either for reasons of downstream processing or for repeated use in order to reduce the specific catalyst costs per kg of product produced. A very simple method is the use of membrane filtration. Because of the increasing number of membranes from different materials (polymers, metal or ceramics) this is an attractive alternative. Whereas for whole cells microfiltration or centrifugation can be applied, for the recovery of soluble enzymes ultrafiltration membranes have to be used120-221. Often immobilization on a support is chosen to increase the catalyst s stability as well as to facilitate its recovery. The main advantages of immobilization are ... 

Howell, J. A., Field, R. W. and Wu, D., Yeast cell microfiltration Flux enhancement in baffled and pulsatile flow systems , J. Membrane ScL, 80, 59 (1993)... 

In continuous ajfinity recycle extraction (CARE) operations, the adsorbent beads are added directly to the cell homogenate and the mixture is fed to a microfiltration unit. The beads loaded with... 

S-layer ultrafiltration membranes (SUMs) are isoporous structures with very sharp molecular exclusion limits (see Section III.B). SUMs were manufactured by depositing S-layer-carrying cell wall fragments of B. sphaericus CCM 2120 on commercial microfiltration membranes with a pore size up to 1 pm in a pressure-dependent process [73]. Mechanical and chemical resistance of these composite structures could be improved by introducing inter- and intramolecular covalent linkages between the individual S-layer subunits. The uni-... 

The microalgae are cultured in bioreactors under solar or artiflcial light in the presence of carbon dioxide and salts. The bioreactors may be closed systems made of polyethylene sleeves rather than open pools. Optimal conditions for pigment production are low to medium light intensity and medium temperatures (20 to 30°C). Pigment extraction is achieved by cell breakage, extraction into water or buffered solution, and centrifugation to separate out the filtrate. The filtrate may then be partly purified and sterilized by microfiltration and spray dried or lyophilized. 

Production of the color involves centrifugal separation of the biomass, cell breakage, and extraction. Use of a salt solution rather than water as an extraction medium increases stability of the color during extraction. Methods for partial exclusion of the polysaccharide from the color extract in order to enhance resolubilization of the dried color were developed. These processes include either microfiltration or co-precipitation of the polysaccharide with an added positively charged polysaccha-... 

Downstream Processing Microfiltration plays a significant role in downstream processing of fermentation products in the pharmaceutical and bioprocessing industry. Examples are clarification of fermentation broths, sterile filtration, cell recycle in continuous fermentation, harvesting mammahan cells, cell washing, mycelia recovery, lysate recovery, enzyme purification, vaccines, and so forth. 

Pharmaceutical Removal of suspended matter is a frequent application for MF. Processes may be either clarification, in which the main product is a clarified liquid, or solids recovery. Separating cells or their fragments from broth is the most common application. Clarification of the broth in preparation for product recovery is the usual objective, but the primary goal may be recovery of cells. Cross-flow microfiltration competes w l with centrifugation, conventional filtration by rotary vacuum filter or filter press and decantation. MF delivers a cleaner permeate, an uncontaminated, concentrated cell product... 

Fritsch, J. and Moraru, C. 1. (2007). Development and optimization of a carbon dioxide-aided cold microfiltration process for the physical removal of microorganisms and somatic cells from skim milk. /. Dairy Sci. 91, 3744-3760. 

Giffel, M. C. T. and Horst, H. C. V. D. (2004). Comparison between bactofugation and microfiltration regarding efficiency of somatic cell and bacteria removal. Bull. Int. Dairy Fed. 389, 49-53. 

Figure 5 Five-channel enzyme sensor for the simultaneous determination of glucose, lactate, glutamate, glutamine, and ammonium. MFM, microfiltration module WV, valves P, pumps DC, dialysis cell B, blank reactors MC, reactor D, biosensor flow cell. (Adapted with permission from Ref. 34.)...

Upon completion of the homogenization step, cellular debris and any remaining intact cells can be removed by centrifugation or by microfiltration. As mentioned previously, these techniques are also used to remove whole cells from the medium during the initial stages of extracellular protein purification. 

As discussed previously, the technique of microfiltration is effectively utilized to remove whole cells or cell debris from solution. Membrane filters employed in the microfiltration process generally have pore diameters ranging from 0.1 to 10 pm. Such pores, while retaining whole cells and large particulate matter, fail to retain most macromolecular components, such as proteins. In the case of ultrafiltration membranes, pore diameters normally range from 1 to 20 nm. These pores are sufficiently small to retain proteins of low molecular mass. Ultrafiltration membranes with molecular mass cut-off points ranging from 1 to 300 kDa are commercially available. Membranes with molecular mass cut-off points of 3,10, 30, 50, and 100 kDa are most commonly used. 

A useful microfiltration apparatus, which prevents re-introduction of dust into filtered solution through contact with laboratory air, has been described by Levine et al. so and is shown in Fig. 12. The liquid is placed in the upper chamber and the lid closed. Filtration into the LS cell and recycling back into the upper chamber are carried out and repeated several times by manipulation of the stopcocks. Finally the filled cell is sealed and placed in the LS photometer. A rather similar device has been reported by Casassa and Berry32. ... 

Fig. 12. Microfiltration apparatus 0 a, b, c and d denote respectively the upper vacuum stopcock, the lower vacuum stopcock, the cell stopcock and the recycling stopcock...

Microfiltration Pressure gradient 10-0.1 p,m Small particles, large colloids, microbial cells... 

Microfiltration (MF) pressure difference filtration of cell suspensions blood plasma recovery... 

Generally, a distinction can be made between membrane bioreactors based on cells performing a desired conversion and processes based on enzymes. In ceU-based processes, bacteria, plant and mammalian cells are used for the production of (fine) chemicals, pharmaceuticals and food additives or for the treatment of waste streams. Enzyme-based membrane bioreactors are typically used for the degradation of natural polymeric materials Hke starch, cellulose or proteins or for the resolution of optically active components in the pharmaceutical, agrochemical, food and chemical industry [50, 51]. In general, only ultrafiltration (UF) or microfiltration (MF)-based processes have been reported and little is known on the application of reverse osmosis (RO) or nanofiltration (NF) in membrane bioreactors. Additionally, membrane contactor systems have been developed, based on micro-porous polyolefin or teflon membranes [52-55]. 

Maiorella B, Dorin G, Carion A, Harano D. Crossflow microfiltration of animal cells. Biotechnol Bioeng 1990 37 121-126. 

Blood cells are separated from blood (hematocrit 40%) by microfiltration, using hollow-fiber membranes with an inside diameter of 300 pm and a length of 20 cm. The average flow rate of blood is 5.5 cm s T Estimate the filtrate flux. 

The cells and cell lysates (fragments of disrupted cells) can be separated from the soluble components by using microfiltration (Chapter 8) with membranes. This separation method offers following advantages ... 

The separation of cell debris (fragments of cell walls) and organelles from a cell homogenate by centrifugation may often be difficult, essentially because the densities of these are close to that of the solution, which may be highly viscous. Separation by microfiltration represents a possible alternative approach in such a case. 

When planning an industrial-scale bioprocess, the main requirement is to scale up each of the process steps. As the principles of the unit operations used in these downstream processes have been outlined in previous chapters, at this point we discuss only examples of practical applications and scaling-up methods of two unit operations that are frequently used in downstream processes (i) cell separation by filtration and microfiltration and (ii) chromatography for fine purification of the target products. 

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