1、Why? What is the most expensive or dearest substance in the world? Update gold price = $1103/ounce=$35.45/g= $ 1.32 million per carats= $ 1.32 million per carats $6.6 million per gram $6.6 million per gramAbout 682 times!It is difficult to separate them:(2) Complex components （成分復雜）(1) desired produ

2、cts in dilute suspension（目標產物濃度低）(3) Biological products require high quality (生物產品高質量)： * purity（純度） * sanitation（衛生） * biological activity（生物活性）How to solve it?What is Bioseparations? (1) A collective term Includes all the steps or methods required in order to recover useful biochemicals or biopro

3、ducts from any kind of industrial processes. ( (所有從任何工業過程中回收有用的生物化學品或生物產品的所有從任何工業過程中回收有用的生物化學品或生物產品的方法、步驟方法、步驟) ) Biochemicals or Bioproducts chemical substances or combinations of chemical substances that are made by living things range from methanol (甲醇) to whole cells. Extracted from whole plants

4、 and animals (提取) synthesized in bioreactors containing cells or enzymes (生物合成)What is Bioseparations(2) Downstream processing for biotechnology What is BioseparationsWhat is biotechnology? What is BioseparationsThe application means the use of carefully cultured microorganisms, animal cells and pla

5、nt cells to produce products useful to humans. （即指利用培養微生物、動植物細胞來生產對人有用的產品。） What is Bioseparations What is BioseparationsAbove all: Biosepartions is a fraction of biotechnology; It consists of a series of steps (or processes);c. Its objective is to obtain ( recover) useful biochemicals or bioproduct

6、s What is BioseparationsUpstream ProcessingbiosynthesisBioseparation Downstreamprocessingdownstream processWhat is BioseparationsBioseparationBioproductsVariety ( structures and sizes )Lability ( temperature, pH, concentration) More unit operationsWhat is BioseparationsThe characteristics of Biosepa

7、rationThe characteristics of BioseparationAmino Acid and their AbbreviationsThe characteristics of BioseparationSmall molecules Primary metabolitesSecondary metabolitesThe characteristics of BioseparationProteinsThe characteristics of BioseparationThe characteristics of BioseparationProteins腺嘌呤鳥嘌呤胞嘧

8、啶尿嘧啶胸腺嘧啶The characteristics of BioseparationThe characteristics of BioseparationThe characteristics of Bioseparation low concentration of the targets (目標產物低濃度目標產物低濃度)The concentration of the targets is very low in fermentation liquor. For example, penicillin: 4.2%, gentamycin(慶大霉素慶大霉素): 0.2%. Moreov

9、er, the concentration of the targets in some animals tissue culture is only about 550 g/ml. Biological (or chemical) activity (活性活性) Bioproducts are sold for their biological (or chemical) activity:Methanol: solvent activity; ethanol: neurological activity; penicillin: antibacterial activity; Taxol

10、(紅紅豆杉醇豆杉醇): anticancer activity, and so on. The characteristics of BioseparationThe characteristics of Bioseparation Fermentation broth being a complicated mixture (complex components) (發酵液成分復雜發酵液成分復雜)The fermentation broth is the complicated mixture of many components( soluble or insoluble), not on

11、ly including macromolecules such as proteins, nucleic acid, polysaccharides but also including intact organisms, fragmented mycelia, other insoluble fractions of medium ingredients, residual substrates, etc. The characteristics of Bioseparation Poor stability ( (穩定性差穩定性差) ) Due to chemical degradati

12、on or microbial degradation, the products can be inactivated by temperature, pH or various enzymes. High purity and strict sanitation criterion Almost all products are used as medicine, biological reagents or food, their purity must meet the need of pharmacopoeia (藥典藥典), reagent criterion or food cr

13、iterion etc. All above makes bioseparations much more difficult than chemical separations!The characteristics of Bioseparation The diversity of products spawns (results in) the broad spectrum of separation methods used(2) Characteristics of Bioseparations Broad spectrum of separation methods （產品的多樣性

14、導致分離方法的多樣性）（產品的多樣性導致分離方法的多樣性） most bioseparation methods come from separations of chemicals ( (大多數生物分離方法來源于化學品的分離方法大多數生物分離方法來源于化學品的分離方法) )The characteristics of Bioseparation(2) Characteristics of Bioseparations This table shows that 80% of all the separation methods for conventional chemicals are p

15、ractical in biotechnology. (大約大約80%80%的化工分離方法可應用于生物分離技術的化工分離方法可應用于生物分離技術) The characteristics of Bioseparation These separations are elaborate and expensive Both dilute feed and products with high purity make the separations be elaborate and expensive.More equipment and labor than all other parts of

16、 the process combined are needed.Costing a substantial fraction of the total cost of the final product. This fraction can be as high as 90% or more!The characteristics of Bioseparation Yeast biomass, penicillin G, and certain enzyme processing costs are percentages of the selling prices which are 20

17、%, 20-30% and 60-70% respectively. Products expressed by genetic engineering, their processing costs can be as high as about 8590% of their selling prices. The processing cost of asparaginase (天冬酰胺酶) used as pharmaceuticals is about 85% of its selling prices because of its high purity. The character

18、istics of BioseparationPrice vs Production紅細胞生成素紅細胞生成素長春堿長春堿紅豆杉醇紅豆杉醇洋地黃洋地黃果膠酶果膠酶The characteristics of Bioseparation The recovery and the purification processes must be well conceived and well designed!Important! Organic molecules, $5 - $10 / g Biopolymers, $1 - $5 / g Food & beverage additives,

19、 $20,000 / g How to lower them?（因此必須仔細考慮和設計產品的回收和純化過程）（因此必須仔細考慮和設計產品的回收和純化過程）The characteristics of BioseparationSo, Before designing, what we should ask and answer are:What is the value of the product? ( (產品價值產品價值) )What is an acceptable product quality? ( (產品質量產品質量) )Where is the product and impur

20、ities in each process stream? ( (產物及雜質在加工過程中出現位置產物及雜質在加工過程中出現位置) )The characteristics of BioseparationWhat are the unusual physicochemical properties of the product and the principal impurities? ( (產物及主要雜質獨特的物化性質是什么？產物及主要雜質獨特的物化性質是什么？) )What are the economics of various alternative separations? (各種分

21、離方法的經濟性比較各種分離方法的經濟性比較) Careful consideration of these questions can help us to determine the optimal separating processes.The characteristics of BioseparationImprove the efficiency of bioseperationReduce the steps of operationsReduce the time of operation processReduce costImprove purityKeep activit

22、yThe characteristics of BioseparationAbove all:這些分離過程不這些分離過程不具備特異性，只具備特異性，只是進行初分是進行初分具有產物的高具有產物的高選擇性和雜質選擇性和雜質的去除性的去除性Downstream processes for production of antibioticProduction of recombinant human alpha-1-antitrypsin (抗胰蛋白酶抗胰蛋白酶)Recovery of an extracellular enzymeNutrient mediumFermentationMicroorg

23、anismsSelf-cleaning clarifierCell massSelf-cleaning clarifierResidual solidsConcentratione.g. ultrafiltration or vacuum evaporationPrecipitationFilter pressDryingEnzyme powderLipidSterilizationLiquid enzymeRecovery of intracellular enzymesNutritive mediumFermentationMicroorganismsSelf-cleaning clari

24、fier (cell harvesting)Cell disintegrationDilution (if possible)Continuous liquid-liquid Cross-flow extraction with aqueous two-phase systemsSelf-cleaning clarifier (separation of cell debris)Concentration and enrichmentPurificationConcentration/ precipitationBioreactor fluid with cellsChromatography

25、Cell disruptionCentrifugal extractionDialysis Cleaning pro insulinPrecipitation Dialysis Sulfonation Extraction Precipitation Chromatography Cell debrissupernatantB-ChainA-ChainRecovery and purification of human insulinBioreactor fluid pretreatmentSterile ultrafiltrationFiltration or centrifugationT

26、wo-stage ultrafiltrationUltrafiltrationPrecipitation Polishing ultrafiltrationRotary filter Formulation DryingFormulation CellsFilter aidprecoatLiquids or solidsCool to 4-5Filter aidLiquid productSolid productDownstream processing for production of liquid or solid proteaseBioreactor fluid with cellsFiltration Extraction pH2-3Back extractionFiltration Crystallization Ion exchangecentrifugationDrying Kerosene and tertiary amineWater