A process of separating electrically

charged particles in solution by passing

an electric current through the solution

Fluid that is separated from clotted blood

Similar in composition to plasma but

lacks fibrinogen and other
substances that are used in
coagulation process
Used an electric field to separate the
proteins in the blood serum into groups of
similar size, shape and charge.

Particularly used to determine

whether the humoral immunity
function normally or not.
SPE is performed to:
• Screen for disease such as multiple
myeloma, Waldenstrom’s
macroglobulinemia and amyloidosis
• Find the causes of
hypogammaglobulinemia (eg:Chronic
Lymphocytic Leukaemia)

Waldenstrom’s Kidney amyloidosis

Multiple myeloma
macroglobulinemia
Based on the heterogeneity of charges of the protein
molecules

Overall charge of protein molecule is determined by

pH of its solvent

pH at which the positive

and negative charges of
a given protein balance
is known as isoelectric
point (pI)
NEGATIVE
POSITIVE
The rate of migration of protein is dependent on:

• The net electrical charge of the molecule

• The size and shape of the molecule
• The electric field strength
• The nature of the supporting medium
• The temperature of operation

In brief, electrophoretic mobility is directly

proportional to
the net charge and inversely proportional to size of
the
molecule and the viscosity of the electrophoresis
medium
 -has lower affinity
for
- speed of separation
protein
(20 minutes to 1
hour) - native clarity after
drying
- ability to store the allow excellent
transparent densitometric
membranes for long examination
periods
Used for electrophoretic separation of proteins in
human serum, cerebrospinal fluid and urine

It separates the human serum into five distinct,

well-resolved zones
Dry Dry

Dry

Dry
The pattern of the
protein may then be
visually interpreted or
quantitated by
densitometer
ALBUMIN GLOBULIN

ALPHA (α) BETA (β) GAMMA(γ)

• The largest protein
component of human
serum
• It functions:
 To keep the blood from leaking out of
blood vessel
 Carry some medicine & other
substances
• Moving toward the negative
portion of the gel
• Involve of α1 & α2

components • α1 is comprised of :

 α1-antitrypsin
 Thyroid binding globulin

 Trancortin

- HDL also include in this fraction

• α2 consist of:

 Ceruloplasmin

 α2-macroglobulin
• Has 2 peaks:
 β1
β
2

• Function:
• Help carry substances, e.g:
iron
• Fight infection
• Beta1

- Composed mostly of transferrin

• Beta2

- Contains β-lipoprotein

- IgA, IgM & sometimes IgG along with

complement also can be identified in this

• Most of clinical
interest focused in
this fraction

• Contains immunoglobulin
(antibodies)
• Function:
Help prevent & fight infection
 HRE was introduced to replaced the traditional 5-
band electrophoretic methods (SPE)

 The reason for this replacement is because SPE is

less sensitive since it can only produces 5 bands

 HRE can produce

12 bands
+ -
Albumin α1 α2 β γ

Albumin α1 α2 β γ
+ -
Albumin α1 α2 β γ
 “β-γ bridging”

+ -
Albumin α1 α2 β γ
+ -
Albumin α1 α2 β γ
+ -
Albumin α1 α2 β γ
+ -
Albumin α1 α2 β γ
However, in a particular disorder, the
serum electrophoretic will show normal
pattern.

Therefore, in these cases, we need to

use the CSF or urine instead of serum.
Detection of Bence
Detection of oligoclonal
Jones Protein in Urine
band in CSF
 Can be used to
differentiate monoclonal
gammopathies and
polyclonal gammopathies
 Can be used to
characterized diseases
based on the patterns of
proteins bands
 If gel not stored in
Patient undergoing
Haemolysed blood will horizontal position,
chemotherapy is not
increased atypical electrophoretic
suitable to take
the globin value patterns will
this test
be produced

Medicines such as
Lipaemic samples are corticosteroids,
Cannot be used for
not recommended for birth control pills&
patient that pregnant
analysis aspirin, will alter the
results
 Serum Protein Electrophoresis is
applicable for screening of
particular disease based on the
patterns of protein bands.
 As for monoclonal
gammopathies patients, further
evaluation such as
immunofixation is required to
determine the specific causes of
the abnormality.