Enzyme Linked Immunosorbent Assay (ELISA) Lab Report (Lab Report Sample)

ENZYME LINKED IMMUNOSORBENT ASSAY (ELISA) LAB REPORT
Name of Class
Student’s Name
Professor’s Name
Enzyme Linked Immunosorbent Assay (ELISA) Lab Report
Nhat My Vu - 000952223
Introduction
Enzyme 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 determine 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 biomedical 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)
Method
Referred to the Practical Handbook Protocol CITATION Dye \l 1033 (Dyer & Pecorino, n.d.)
Result
1 Standard Calibration Data
Concentration

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)
34290012255500
Figure 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 1
21/10/16

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 2
9/11/16

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 3
11/11/16

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

18.5974


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.
4286251651100
B.
4572004508500
5048258572500C.
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 control 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...