DiscussionSerum protein electrophoresis has proven to bean effective and efficient method to separate different protein components inthe serum. Proteins are substances made of smallchemicals called amino acids and play a number of key roles in order to keepeveryone alive. Too much or too little protein can cause problems. (4). Ingeneral, the higher the concentration of serum in a sample, the greater concentrationof protein there should be.
Proteins carry a negative or positive charge. The electricalcurrent is what separates the serum proteins into the five major groupsaccording to their size, shape and net charge. Albumin is heavily negatively chargedso therefore it migrates furthest to the anode which is visible from the gel. Likewise,gamma globulins are positively charged so therefore it moves towards thecathode, negative proportion on the agarose gel. The migration of the proteinsalso depends on the properties of the agarose gel and its strength of current.
The most abundant protein in the blood is Albumin as its showingthe darkest and thickest bands meaning greater level of Albumin in the blood. Albuminis made by the liver that keeps fluid from leaking out of the blood vessels andregulates the oncotic pressure. (6) However,in the test samples, the darkest band is at samples 6 which must have a highestconcentration of gamma globulins compared to standard samples and the remaindertest samples. Gamma globulins provide short-term protection and reduce severityof certain diseases and make up the largest proportion of immunoglobulins. Thereason for the Gamma globulin bands being blurry and wide is due to multipledifferent types of immunoglobulins being present in the gamma field.
Mixture ofshapes and sizes so therefore each one runs differently on the gel. (7)SPE is a simple and rapidmethod of separating proteins in the serum with immediate results.Electrophoresis is proven to be extremely accurate giving reliable results andis low in cost to use approximately around $2.
60 per gel analysis. (8)The weaknesses of this method include; alimited sample analysis can be done so only a known sample can be analysed andnot very specific or into greater depth of unknown samples. Also with SPE astarting sample is required so a sample must be prepared in advance in order tomeasure what components are in the sample. So, a large tissue sample isrequired in order to run the analysis and almost impossible to run on a singlecell sample. The limitation is that electrophoresis of proteins can onlyidentify main big groups not into depth of more smaller components.
(9) In the future, a better method suggestedwould be Western blotting which is an analytical technique used to pinpoint a specific proteinin more detail. It’s a longer process but is more accurate and the specificityis greater. The western blot method is more favourable as the sensitivity isvery good. Can detect as small as 0.
1ng of protein in a sample whichwould make a perfect diagnostic tool than can detect tiny immunogenic responsesfrom a virus or bacteria in the sample of a patient.