Abualqasim karam Abualqasim

22101164
Applied Health Sciences Faculty of
r the Technology medical laboratories Program Unde
MLT321 supe
n of rvisio
Dr Adel A. Elazab Elged

Assignment
Competitive immunoassays
The targeted analyte of interest (antigen) competes with a constant
added amount of labelled similar antigen for a limited amount of
specific antibody binding sites. The amount of labelled antigens bound
with the restricted antibodies is Inversely proportional to the amount
of analyte of interest present in the specimen, meaning that as the
amount of the analyte in the sample Increases, the detectable signal
.decreases
the assays are highly sensitive and are able to produce a large signal
from a small amount of analyte. The sensitivity of each assay is related
.any inversely proportional to the affinity constant of the antibody used
Competitive immunoassays can be classified further as
:Simultaneous addition
.Where all components are added at once
:Sequential
where the sample is incubated with the antibody before the labelled
.analogue is added. This assists to improve the sensitivity of the test
 Non-competitive immunoassays
or immunometric immunoassays utilize an excess of labelled antibody ,
toward the analyte of interest (the antigen). “Sandwich” immunoassays
incorporate two separate antibodies, one labelled, and one un-labelled
and fixed to a solid support which is used to extract the analyte of
interest from the sample. The second labelled antibody is added and
binds to a separate site on the bided analytes- resulting in an antibody
“sandwich” with the analyte positioned in the middle of the two
antibodies. The amount of analyte is able to be determined through
measuring detectable signal from the labelled antibody, whereby the
amount of bound labelled antibody is directly proportional to the
amount of analyte present in the specimen
Non-competitive immunoassays are highly specific and sensitive, being
able to theoretically detect just one molecule of a specified antigen.
Due to the excess amount of antibody present, non-competitive
immunoassays are significantly faster and less prone to influence by
substances or conditions affecting the reaction equilibrium compared
.to competitive immunoassays

Table summarizing the key differences between competitive and non-competitive immunoassays:
Non-competitive immunoassay Competitive immunoassay Feature 1

Excess of labeled antibodies captures analyte, Competition between labeled analog and unlabeled
resulting in a sandwich complex analyte for antibody binding sites 2
Principle

Increases as analyte concentration increases Decreases as analyte concentration increases Signal 3

Generally more sensitive than competitive

assays Generally less sensitive than non-competitive assays 4
Sensitivity

Often used for analytes with low Widely used for a variety of analytes, including
concentrations in samples hormones, drugs, and toxins Applications 5
Competitive immunoassay

 Thyroid-stimulating hormone (TSH) is a hormone that is

produced by the pituitary gland and stimulates the thyroid
gland to produce thyroid hormones. The cobas e411 TSH
assay is a competitive immunoassay that uses a labeled
 analog of TSH to compete with the unlabeled TSH in the
sample for binding to antibody binding sites.
 Thyroxine (T4) is a hormone that is produced by the thyroid
gland. The cobas e411 T4 assay is a competitive
immunoassay that uses a labeled analog of T4 to compete
with the unlabeled T4 in the sample for binding to antibody
binding sites.

The assays are sensitive and specific, and its can be used to
diagnose and monitor thyroid disorders.

Non-competitive immunoassay

 C-reactive protein (CRP) is a protein that is produced by

the liver in response to inflammation. The cobas e411 CRP
assay is a non-competitive immunoassay that uses an
excess of labeled antibodies to capture the CRP in the
sample.
 The assay is sensitive and specific, and it can be used to
assess the risk of cardiovascular disease

 Myoglobin: Myoglobin, a protein found in muscle cells,

serves as an oxygen storage protein. The cobas e411
myoglobin assay utilizes a non-competitive immunoassay
method, employing an excess of labeled antibodies to
capture the myoglobinpresent in the sample. The assay's
sensitivity and specificity make it useful for diagnosing and
monitoring muscle damage.

Noncompetitive (immunometric) immunoassay An

immunoassay in which the patient's analyte Is allowed to bind
.with an excess amount of labeled reagent
Competitive immunoassay An Immunoassay in which the
patient's unlabeled analyte competes with a constant amount of
.labeled analyte for a limited amount of reagent
  What are the criteria for acceptance of the results of the linearity study?
C B A

Acceptable value Description Criterion 1

Correlation
r > 0.99 Strength of linear relationship coefficient (r) 2

Randomly distributed within acceptable limits Deviations from fitted line Residuals 3

Measured
Within acceptance limits Values at upper and lower limits of reportable range responses 4

Reference
Thank you