Immunology Lab Report

Michael Ghermezi and Aaron Whitby

Email: mikeghermezi@gmail.com, aaronwhitby.ucla.edu

ID: 92555067886 (MG), 917701799 (AW)







































Introduction: In this lab, we conducted an Enzyme Linked Immunosorbent Assay
(ELISA) to determine immunoglobulin levels. Commonly, the ELISA test has been
used as a diagnostic tool in medicine and pathology. The process is based on specific
binding of an antigen via specific antibodies. Antigens from a sample are attached to
a surface (i.e. well/microtiter plates) Then, a specific antibody is applied to bind to
the antigen. This antibody is linked to an enzyme, and lastly a substance containing
the substrate is added to the wells. The reaction produces a detectable colar change
in the substrate.
The ELISA test has many applications, such as identification and
quantification of substances in tissues and body liquids, specifically hormones,
drugs, markers, etc. Additionally, ELISA can be utilized to quantify antibodies in
body tissues and fluids. In this lab, we calculated the HIgG from the sample patient
and the primary antibody was linked to the HRP enzyme.

Lab Objective:







Calculations:
After averaging the values for rows one and two and subtrating the average of the
blank plates we came up with the following data

Absor
bance
450n
m
(OD)
1.1361
667
1.183
1667
1.19
0666
7
1.09
266
67
1.0
126
667
0.838
6667
0.636
1667
0.444
6667
0.3206
667
0.24416
67
0.18316
67
0.17266
67
Conce
ntrati
on
3 1.5 0.75 0.37
5
0.1
875
0.093
75
0.046
875
0.023
4375
0.0117
1875
0.00585
9375
0.00292
9688
0.00146
4844

We plotted the absorbance vs concentration (Log). We then took the most linear
data points in the center and had excel calculate our equation.

y = 3.2314x + 0.449








1.1361667
1.1831667
1.1906667
1.0926667
1.0126667
0.8386667
0.6361667
0.4446667
0.3206667
0.2441667
0.1831667
0.1726667
y = 3.2314x + 0.449
R = 0.9066
0
0.2
0.4
0.6
0.8
1
1.2
1.4
0.001 0.01 0.1 1 10
A
b
s
o
r
b
a
n
c
e

(
O
D
)

Concentration (mg/ml) Log
The averages of our absorbance for the unkown samples are as follows as wella s
the calculated concentrations:
Unkown 1 Unknown 2 Unknown 3
1.338 .774 .076
Calculated
concentration X
100 to correct
for dilution
27.5 10.1 -11.5
Diagnosis Hyper HigG Normal HigG No HigG