2. Outline
I. Introduction
I. Protein Separation
Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE)
II. Protein Identification
Western Blotting
Enzyme-linked Immunosorbent Assay (ELISA)
Mass Spectrometry
3. Introduction
Proteins in the body form the structure of cells; make up enzymes and hormones that
regulate bodily functions; transport analytes, nutrients, and drugs throughout the body;
remove metabolic waste; and provide defenses against infection.
Among the classes of molecular analytes found inside the body, proteins provide a
unique combination of broad clinical significance and accessibility.
The analysis of proteins and clinical diagnostic tests for proteins are widely accepted in
medical technology
4. Sodium Dodecyl
Sulphate
Polyacrylamide Gel
Electrophoresis (SDS-
PAGE)
A technique used for separating the proteins based
on their molecular weight.
Sodium dodecyl sulphate and polyacrylamide help
to eradicate the influence of structure and charge of
the proteins, and the proteins are separated based
on the length of the polypeptide chain.
https://youtu.be/MILiO1XnuqQ
5. Western Blotting
Was first coined by Dr. Burnette in 1981 after the
eponymous southern blot for DNA and consequent
coinage of the northern blot in 1977 for RNA
WB selects for an individual protein amongst a
potentially significant milieu via leveraging the
specificity of antigen (Ag)-antibody (Ab) binding.
https://youtu.be/qT_AqI-MKJc
6. Enzyme-linked
Immunosorbent
Assay (ELISA)
Is a labeled immunoassay that is considered the
gold standard of immunoassays. This immunological
test is very sensitive and is used to detect and
quantify substances, including antibodies, antigens,
proteins, glycoproteins, and hormones. The
detection of these products is accomplished by
complexing antibodies and antigens to produce a
measurable result.
ELISA testing is used to diagnose HIV infection,
pregnancy tests, and blood typing, among others.
https://youtu.be/RRbuz3VQ100
7. Mass
Spectrometry
Is a mainstream chemical analysis technique in the
twenty-first century. It has contributed to numerous
discoveries in chemistry, physics and biochemistry.
Hundreds of research laboratories scattered all over
the world use MS every day to investigate
fundamental phenomena on the molecular level.
Commonly regarded as an instrumental technique
for separation of electrically charged species in the
gas phase: the charged species (ions) are produced
in the ion source (in some cases, the ion source also
assists the transfer of solid-phase; or liquid-phase
analytes into the gas phase); the gas-phase ions
subsequently are transferred into the mass analyser,
which in turn sorts the ions—in space or time—
according to the massto-charge ratios (m/z); finally,
the separated ions are detected by an ion detector in
the space or time domain.
https://youtu.be/JPpKL1uzE0I
