The Intensity Of Agglutination

The Intensity Of AgglutinationAntibodies ar proteins produced during bodies defence against foreign antigens and they atomic number 18 driven from plasma cells. In the emergence of an immune solution B lymphocytes initiate the production of immunoglobulin M antibody. In comparison to other immunoglobulins immunoglobulin M is the braggyst and earliest antibody available in response to an antigen (Bailey Johnson, 2006). The large structure of this antibody is because it consists of an humanityitarianal domain in its constant sweep (Overfield et al, 2007). This antibody has a polymeric structure it consists of sober and send chains. The binding amid two heavy chains or between heavy and light chains is facilitated via the disulphide mystify. immunoglobulin M antibody has a pentameric structure consisting of five subunits. These subunits are join together via a disulphide bond which occurs between the Fc region and the intersubunit, interasubunit- J chain. twain fab antig en binding sites are available on each IgM monomer and since IgM has a pentameric structure ten Fab antigen binding sites are available that gouge potentially act with ten antigens (Overfield et al, 2007)(Khurana, 2006). The initial de sign of the zodiacing of this practical was to discover if rubicund crease cell antigens provoke interact with IgM anti-D (Rh) antibody and weather as a consequence of this interaction agglutinating activity occurs. The second aim was to discover weather dithiothreitol (DTT) reducing cistrons is capable of fixing the structure of IgM antibody at different niggardliness accordingly touching the level of agglutinating activity and finally to discover if indirect anti-IgM antibody is capable of facilitating agglutinating activity. The large and pentameric structure of IgM antibody can potentiate the possibility of its interaction with red prodigal cell antigens resulting in constitution of agglutination.Material MethodFor instruction manual on how to conduct the experiment with the relevant materials used please push to the practical schedule. The slow-wittednesss of DTT added to the nine tubes where as following (0.001, 0.002, 0.003, 0.004, 0.005, 0.006, 0.007, 0.008, 0.009 0.01).ResultsTable 1 The preceding(prenominal) table illustrates the number of tubes denominate from 1-10 and the concentrations of DTT in (Mol/L). As illustrated in the preceding(prenominal) table the control tube which is tube 1 lacked DTT maculation tubes which were numbered as (2, 3, 4, 5, 6 7) consisted of different concentrations of DTT as shown here (0.001, 0.002, 0.003, 0.004, 0.005, 0.006 0.007). gibe to the first contemplation results tubes numbered 1-7 expressed signs of agglutination as indicated by a positive sign (+). Instead tubes numbered (8, 9 10) which had the following DTT concentrations (0.007, 0.008 0.009) expressed no indications of agglutination hence they were marked as negative (-). Due to time demarcations re sults for the second antibody labelling could not be obtained.DiscussionThe inspiration of agglutination in these tubes depended on the concentration of DTT. The control tube which is tube 1 is DTT deficient which is accompanied with agglutination. Tubes labelled 2-7 express different concentrations of DTT starting from the lowest hence escalating slowly. In these tubes agglutination is still observed since the performance of DTT is still not strong copious to break the bonds expressed in IgM antibody while as the concentration of DTT escalates further in tubes 8-10 agglutination is not evidenced. DTT is a reducing agent capable of mediating intersubunit and interasubunit-J chain cleavage hence facilitating IgM subunit (22) synthesis (Kownatzki Drescher, 1973). As the concentration of DTT escalates its capability to break these bonds with greater intensity increases as seen in tubes 8-10 jumper cable to greater IgM subunit formation (22) and lessens the possibility of antigen a ntibody interaction hence lack of agglutination. In addition DTT affects the structure of IgM heavy and light chains by preventing them from efflorescence and causes this chain too separate accordingly atomic number 82 to agglutination deficiency. A continuous raise in DTT concentration as evidenced in tubes 8-10 causes a decline the probability of disulphide bonds from resuming their attend in IgM antibody (Valetti Sitia, 1994). agree to the study conducted by (Marrodan et al, 2001 Morris et al, 1974) DTT reducing agent restrains agglutination from occurring by facilitating the disulphide bond located in the IgM antibody to break. In addition the 19 S IgM antibody is cleaved by DTT into a 7S subunit. The 7S antibody subunits are rendered incapable of maintaining IgM antibodys function and therefore wont be able to interact with red blood cell antigens leading to lack of agglutination (Knight, 1978).Due to time limitation for the experiment results for the second antibody labe lling could not be obtained. According to (Overfield et al, 2007) the lacking agglutination as a consequence of DTT effect can be change by adding anti-IgM antibody hence signs of agglutination will appear but the extent of agglutination will depend on whether the IgM antibody subunits have well-kept their ability to bind to red blood cells antigen or due to senior high level of DTT concentration they have been completely deformed.According to the study conducted by (Emmerich et al, 2006) IgM antibody can be used in the diagnosis of capital of Tibet virus infection which is highly predominant in Western African patients. This diagnosis is achieved via victimisation reverse enzyme immunoassay (ELISA) technique to identify anti-Lassa IgM antibody. The result of this study implemented that via using reverse ELISA in 20 patients with sign of fever high level of anti-Lassa IgM antibody was diagnosed indicating the carriage of the Lassa virus. In a study conducted by (Varsano et al, 1995) the strawman of IgM antibody against respiratory syncytial virus antigen (RSV) was examined in 145 patients via using the ELISA technique. According to the result of this study ELISA-IgM antibody chance uponion is a highly efficient order in the diagnosis of RSV at early stage of the disease. In other(prenominal) study by (Tsuda et al, 2001) polymerase chain reaction (PCR) technique was used to detect for the presence of IgM antibody against TT virus (TTV) in the diagnosis of human circovirus. The result of this experiment suggests that healthy volunteers were defective of anti-TTV IgM antibody whereas infected individuals showed signs of its presence suggesting that this method is beneficial for diagnosis purposes of human circovirus.Immunoglobulin cleavage can be triggered via the action of different enzymes or chemicals. Papain is an enzyme that cleaves IgG antibody into three segments of FC, heavy and light chains. Furthermore IgM antibody can be cleaved by pepsin enzy me all into an antibody that weights less accompanied with FC fragments (Rudmann, 2005)(Svehag et al, 1969). Protease enzyme is driven from Neisseria gonorrhoeae bacteria capable of cleaving immunoglobulin A antibody (Pouedras et al, 1992). According to (Akesson et al, 2006) streptococcus pyogenes bacteria is responsible in mediating diseases such as gotonsillitis, septicaemia and it intervenes its action by causing IgG antibody cleavage via using an enzyme called Ides. The action of this virus is to insure that the antibody is unavailable to destroy the bacteria. Furthermore trypsin is another enzyme capable of cleaving IgM antibody at temperature above 50 C leading to different FC fragment synthesis (Andrew et al, 1970).ConclusionNormally red blood cell antigens are capable of interacting with IgM antibody resulting in agglutination while in the presence of DTT reducing agent this binding is inhibited leading to lack of agglutination. The extent of this inhibition will depend on the concentration of DTT and the extend of IgM J chain, interchain intrachain cleavage via DTT. The greater the concentration of DTT the stronger its effect is on this chain which lessens the likelihood of this chain regaining their binding condenser hence their ability to regain antigen binding activity. The concept of antigen antibody binding can be used for the diagnosis purpose of many diseases.