University:
Swansea University
Course:
PM134 | Microbiology
Academic year:

2023
Views:

228

Pages:

3
Author:

Amal J.

spaceaimAffinitymatriseanzymeBound enzyme - wash free ofaffinity elutioncontamerating proteins(with as )(OR)+dialysisRestore optimum conditionsPurified erzymeMatrix Properties :-possess suitable and sufficient chemical groups to whichthe ligard may be covalently coupled and be stable anduthe conditions of the attachmentbe stable during binding of the macomolecule and itsSubsequent elution.interact only - weekly with other maaomolecule to miringenon Specific adsorptionexhibit good flow propertiesones areParticles (uniform, sphere, rigid) are used. Most commonCrosslisked declar againse, polyaerylamids polymethouglatepolystere cellulose & porous glass and Slica. FPLLFPLC: Fast performance liquid cheumatography:-A fast A type of chromatography where the Solvent velocityis controlled by pumps The pumps controll the constantflow rate of the Solverts.The Students are accessed through tubing from an outsiderescivoir.The flow rate of the Solvest is set through comp inputand controlled by pumps.There cue various columns in log chromatography dependingon the type of Separation preferred Each Column containsa small diameter packing material. The column is a largeCmm id) tube containing (u) small particles (gel beads)known as stationary phase.The cheomatographic bed is composed by the gel beadsalone when they are inside the column-The sample is introduced into the injector and thencarried into the column by the following solventOnce in the Coloumn, the sample mixture separatesas a result of different Components adhering to ordiffusing into the gel.As the Solvents is forced intoth chromatographic bedby the flow rate, the samples Separates into various Lones ofSample conc. These zonesbands.Biomolecules have various characturities such as Molecularweight electric change and hydrophobicityAs do purification of the object is usually achieved byusing a combination of chromatographic methods: HPLCNot limited by sample volativity or thermal stabilityRoom temperature analysaEase of sample recovery.Sample injection loopcludersamplecollectorSolventres ervoirHigh previuepumpRecorder anddata systemColumns ofconventional columns are usedGenerally made of stainless Steel Ey all manufactured sothat they can withstard . pressure of up to SOMPa.3- 25 cm long 4:6 nmm intunal diameterFlow rate 13.m3 min !Microbore / open tubular columninternal diameter1-2 mm,2550 cm long, They can sustain flow rade of520 mm 3min.'.Advantages of microbore over convertional :-Reduced cluent consumption due to the S lower flowratesK ideal for interfacing with a mars spec due tothereduced flow ratered Sensitivity due to the higher Conc of analyte thatCan be used