Enzyme Linked Immunosorbent Assay (ELISA) Lab Report - 2200 Words

Enzyme Linked Immunosorbent Assay (ELISA) Lab Report (Lab Report Sample) Content: ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) LAB REPORTName of ClassStudents NameProfessors NameEnzyme Linked Immunosorbent Assay (ELISA) Lab ReportNhat My Vu - 000952223IntroductionEnzyme Linked Immunosorbent Assay abbreviated as ELISA is a quantitative analytical technique used primarily in immunology. The technique is carried out to detect and measure or estimate the quantities of antibodies using ligand conjugated to an enzyme that makes a substrate change its color CITATION Del11 \l 1033 (Delves, et al., 2011).This test can be used to determine if you have antibodies related to certain infectious conditions. More often than not, an Elisa test serves as a diagnosis to HIV/AIDS, Lyme disease, pernicious anemia and syphilis, among many other infections. CITATION Kin15 \l 1033 (Kinman, 2015). An ELISA test serves as a screening tool before in-depth tests are ordered. Supposing one shows the signs and symptoms of a given condition, an ELISA test is administered to dete rmine the presence or absence of the condition. CITATION Kin15 \l 1033 (Kinman, 2015) The purpose of this lab is to determine the presence of serum antibodies that target the HIV antigen. There are four different types of ELISA techniques, which include are direct ELISA, indirect ELISA, capture ELISA (sandwich), and competitive ELISA. The two main ELISA techniques that detect the amount of antigens or antibodies are direct ELISA and indirect ELISA. Direct ELISA involves the surface of the well of the strip covered directly with antibody or antigen, in other words the wells coated directly with primary antibodyCITATION Ayd15 \l 1033 (Aydin, 2015). Indirect ELISA involves the attachment of the antigen to the polystyrene plate, but in this case, the primary antibody is not labeled. An enzyme-conjugated secondary antibody is then added. This format is used most often to detect specific antibodies fin sera. Sandwich ELISA involves the attachment of a capture antibody to the polystyrene plate. Samples containing known or unknown antigen are then added in a matrix or buffer that will minimize attachment to the solid phase. An enzyme-labelled antibody is then added for detection. The competitive ELISA involves the simultaneous addition of competing antibodies or proteins. The decrease in signal of samples where the second antibody or protein is added gives a highly specific result. CITATION Eli \l 1033 (Anon., n.d.) This experiment was using indirect ELISA technique to test the two unknown samples given for the specific antibodies that is in the presence of HIV, and to define the amount of antibody in each patient sample CITATION Dye \l 1033 (Dyer Pecorino, n.d.). Moreover, this lab involved a serial dilution, which is a method, used to determine an antibody concentration by measuring the change of the color spectrometically (e.d). The ELISA test is used in many different fields such as for biological purposes, nutrition, biomedical, and pharmaceutical. In biome dical fields, ELISA is used in research and diagnosis laboratories to detect the concentration of different things such as concentration of hormone, peptides, and proteinsCITATION Ayd15 \l 1033 (Aydin, 2015)MethodReferred to the Practical Handbook Protocol CITATION Dye \l 1033 (Dyer Pecorino, n.d.)Result 1 Standard Calibration DataConcentration Group Absorbance Mean Absorbance of Whole Class Data 0 0.058 0.081 1 0.076 0.081 2 0.086 0.111 4 0.069 0.111 8 0.089 0.107 16 0.115 0.128 31 0.306 0.156 63 0.248 0.230 125 0.333 0.285 250 0.381 0.351 500 0.440 0.399 1000 0.366 0.408 Table 1: Individual group absorbance and the whole class absorbance mean, this table used to graph the standard calibration curve. (Data in green field represent the range of concentration data (16ng/ml 500ng/ml) was used for analysis, which is used to calculate the concentration of control samples and unknown samples)34290012255500Figure 1: Standard Calibration Curve of individual group vs absorbance means of the whole class data. An individual group calibration curve compared to the whole class mean curve is slightly different even though the both curve having increasing trend but the individual group curve is fluctuated for a little bit. 1 Each group data of the whole class data set.GROUP Slope Intercept Positive Abs Positive Conc Patient A Abs Patient A Conc Patient B Abs Patient B Conc Group 121/10/16 1 0.0008 0.1159 0.3993 370.0064 0.3630 322.572 0.1790 82.3530 2 0.0005 0.1479 0.4097 509.8701 0.3983 487.797 0.2287 167.3500 3 0.0008 0.1208 0.3843 349.5377 0.4873 477.3941 0.3670 328.0214 4 0.0005 0.1434 0.3327 394.4098 0.3073 341.6240 0.2130 145.0666 5 0.0001 0.3799 0.4227 571.0129 0.4187 517.6560 0.2647 -1536.5838 6 0.0001 0.1374 0.1670 311.8762 0.1283 -96.0835 0.1043 -349.2998 7 0.0006 0.1519 0.3963 406.9195 0.3413 315.3670 0.1830 51.8068 Group 29/11/16 8 0.0007 0.0851 0.4097 496.8844 0.4300 528.0129 0.2600 267.7582 9 0.0005 0.1108 0.3373 491.0946 0.3973 621.1433 0.2993 408. 7305 10 0.0005 0.2027 0.4077 396.7317 0.4010 383.8293 0.2963 181.2623 11 0.0007 0.1611 0.4207 396.8237 0.3937 355.5417 0.2653 159.3250 12 0.0007 0.1308 0.3977 382.4948 0.3720 345.7086 0.2227 131.6802 13 0.0006 0.2033 0.4293 376.6480 0.3147 185.5537 0.3727 282.2118 14 0.0007 0.1824 0.4510 383.7400 0.4453 375.6430 0.2660 119.3947 15 0.0004 0.2433 0.3573 265.9704 0.3113 153.7268 0.2307 -29.3381 16 0.0005 0.2242 0.3850 313.7208 0.3947 332.5776 0.2527 55.5774 17 0.0007 0.1783 0.4967 441.5708 0.4140 326.9104 0.3150 189.5954 Group 311/11/16 1 0.0005 0.2229 0.2873 130.6771 0.2890 134.0599 0.1890 -68.9070 2 0.0006 0.1234 0.4730 563.8919 0.4410 512.2797 0.3187 314.9709 3 0.0002 0.0779 0.0743 -15.1391 0.0557 -93.8930 0.0597 -77.0171 4 0.0002 0.3315 0.6323 1730.381 0.4807 853.1318 0.3280 -19.8683 5 0.0008 0.1156 0.5037 472.8354 0.5047 474.0539 0.2403 151.9824 6 0.0004 0.1592 0.3653 570.6485 0.3063 407.3175 0.1667 20.6754 7 0.0005 0.1804 0.3060 242.2535 0.3060 242.2535 0.1900 18.59 74 8 0.0005 0.1890 0.3287 263.0817 0.3213 249.2650 0.1780 -20.7887 9 0.0006 0.1946 0.3553 283.4944 0.1247 -123.278 0.3490 272.3258 10 0.0006 0.1493 0.0573 -161.497 0.1347 -25.7309 0.3490 350.5530 Table 2: Absorbance and Concentration of Positive Control, Patient A and Patient B Samples. Absorbance was measured by the microplate reader and concentration was calculated by using intercepts and slope from excel from each group in the whole class data set. All the red letters represents the groups that have outliers data, the grey highlight is the actual outliers value.3. Whole class dataset for Positive control, Patient A, and Patient B, also define the outliers from each set.A.4286251651100B.45720045085005048258572500C.Figure 2: (A) Positive control, (B) Patient A, (C) Patient B concentration data from table 2. Graph demonstrated the whole class data of positive control, Patient A and Patient B concentration. Red dots on the graph represent the outliers in the set of positive contro l data. In these three graphs, there are some extreme values that are out of range and can clearly observed through the graph, which indicates the outliers. Outliers can be extremely high or extremely low. 1 Standard deviation, mean, and median for positive control, patient A, and patient B from the whole class Mean Median Standard deviatio...