PBBS502 Lecture3 Blood Proteins

Blood, Plasma and Serum Components
Gabriella Cs-Szabo, Ph.D.

Department of Biochemistry
Gabriella_Cs-Szabo@rush.edu
Resource Material:
Devlin Chapter 3.3
Lecture objectives:
1. Describe the most important blood components together with their functions.
2. Define what serum and plasma are.
3. Describe the sources of serum proteins and the importance of albumin in the blood.
4. Describe the properties of a serum electrophoretogram. Name the major peaks and
list some proteins under each peak.
5. Describe the change in electrophoretic pattern of plasma proteins for a patient with
liver disease, kidney disease, monoclonal gammopathy, infection and
immunosuppression.
Key Words:
Blood, plasma, serum, electrophoresis, serum proteins, albumin, gamma-globulins,
monoclonal gammopathy
BLOOD, PLASMA AND SERUM COMPONENTS

1.
2.
What is blood?
Blood is a major connective tissue that contains water,
dissolved elements and specialized cells.
It is enclosed in the vasculature and moved around by
the heart.
.
What is the function of blood?
A. Transport:
Nutrients
Waste products of metabolism
Gases (dissolved or bound to hemoglobin in RBC)
Hormones
Enzymes
Plasma proteins
Blood cells
B. Maintenance of pH (buffer systems)
C. Regulation of body temperature
D. Elimination of waste products
E. Regulation of body fluid electrolytes
F. First line of defense
Human circulatory system

Free domain
Blood components are exchanged with the environment to fulfill the function of
blood.
Blood receives oxygen from the lungs, nutrients from the gut, takes up waste
products from the periphery, and eliminates carbon dioxide in the lung and waste
products in the kidney.
Gas exchange in alveoli

LadyofHats Public domain
The circulatory system

Public domain
3. What are the major components of blood?

55% of blood is plasma
45% of blood is cells - 99% erythrocytes (RBC)
1% leukocytes (WBC) and thrombocytes (platelets)
Blood components
http://training.seer.cancer.gov/ss_module08_lymph_leuk/images/illu_blood_components.jpg
A. Major functions of the cells:

Erythrocytes: transport of blood gases
Leukocytes: lymphocytes
monocytes
neutrophils
basophils
eosinophils
defense mechanisms (antigens, microbes, parasites), immune response,
inflammatory response
Thrombocytes: blood coagulation
Electron Microscopy Facility at The National

Cancer Institute at Frederick (NCI-Frederick)
From left to right:
erythrocyte, thrombocyte, leukocyte
Courtesy: Department of Histology, Jagiellonian

University Medical College
a erythrocytes; b neutrophil;
c eosinophil; d - lymphocyte
3. What is plasma and serum?

The plasma is the medium in which the blood cells and dissolved components are
transported around the body.
Normal blood plasma is 90-92 % water, in which different molecules are suspended.
Dissolved substances include
electrolytes such as sodium, chlorine, potassium, manganese, and calcium ions
nutrients and waste products
lipoproteins transporting lipids and lipid-soluble molecules
hormones (transported by proteins)
plasma proteins (albumin, globulin, fibrinogen, etc) 98% of total solutes
Human plasma constituents by volume.

Courtesy by Schmin public domain
Cells are separated from the fluid component by centrifugation.
Plasma is gained when anti-coagulants are given to the blood
Serum is gained after coagulation, thus serum is fibrinogen-free plasma (also low in
blood clotting factor levels).
Scheme of a blood sample

after centrifugation
KnuteKnudsen Public domain
Blood samples, right: freshly drawn;

left: treated with EDTA (anticoagulant)
JHeuser Public domain
4. Why do we use blood tests?

To determine specific cell numbers and cell ratio
To determine plasma (serum) composition
Test for a specific component in plasma (serum)
Since proteins are the major non-cellular component of the plasma, the analysis
of proteins may be used to assess certain disease states.
5. What kinds of proteins are there in the blood?
Plasma proteins are of two types:
a. those at high concentrations, specific to blood and with a functional role in blood,
for example albumin, carrier proteins, immune proteins, proteins of blood
coagulation
All plasma proteins, except for the immunoglobulins, are made in the liver.
b. those at low levels, with no functional role in blood.
proteins that are released from the cells because of normal cell turnover
In disease, there may be an increase or decrease in the levels of resident proteins.
a. increase or decrease of the protein levels that are specific for blood (e.g. albumin
or gamma-globulins)
b. increased release of intracellular proteins into the plasma, thus increased levels
of proteins that normally are in a low concentration in blood (e.g. heart enzyme
levels in case of MI).
Figure Lippincott. Increase of protein levels in blood by increased level of cellular release
Plasma proteins can be detected and separated from each other based on their physicochemical characteristics.
Electrophoresis is used
to identify the presence of abnormal proteins,
to identify the absence of normal proteins, and
to determine when different groups of proteins are increased or decreased in serum or
plasma.
6.
Analysis of plasma proteins in diagnoses of certain diseases
In clinical medicine, separation of serum or plasma proteins can be used to classify
proteins and also used to diagnose diseases.
Proteins can be separated based on their net charges. The net charge of the protein
depends on its amino acid composition and the pH of the environment. Proteins in
which ratio of Lys+Arg/ Glu+Asp is greater than 1 are basic and those less than 1 are
acidic proteins.
Electrophoresis of serum or plasma is usually done at pH 8.6, at which the proteins are
negatively charged and move toward the (+) electrode at a rate dependent on the net
charge.
o Major peaks are those of albumin, alpha (alpha1- and alpha2-), beta- and gamma
(gamma1- and gamma2-) globulins.
o Variations in these peaks are indicative of certain diseases, thus can be used in
diagnosis.
o Usually biuret reaction is used to measure total protein content of plasma, and peaks
are given as % total protein.
(FYI: In biuret reaction, copper ions form colored complex with the amide nitrogens
of the peptide bonds of the protein backbone.)
Albumin is a peak that composed of albumin only; the other peaks are mixtures of
different proteins.
Figure 3.20. Devlin Electrophoresis pattern for plasma proteins at pH 8.6.

The plot shows the order of migration along the horizontal axis with proteins of highest
mobility closest to the positive electrode. Area under the peak is proportional to
protein concentration. Different major proteins are running under each peak (except
for the albumin peak).
Examples of plasma proteins:

Albumin: The most abundant plasma protein that makes up 60% of the total protein of
the plasma. Its function is keeping the osmotic pressure of the blood and
transportation of biomolecules. It transports hydrophobic compounds, e.g. bilirubin,
fatty acids, hormones, etc.
Clinical relevance: When albumin level is low, water moves from the arteries to the
interstitial space and cannot return to the veins; resulting in edema. Thus, any
disease that decreases the levels of albumin in the plasma may result in edema.
Examples of hypoalbuminemia (low albumin level in blood):
In liver disease, associated, for example, with hepatitis C infection, alcoholism, etc, the
malfunction will result in diminished production of albumin.
Kwashiorkor disease, protein malnutrition in children, when essential amino acids are
not taken up in the required concentration, albumin will not be synthesized, thus
fluids accumulate especially in the abdomen (ascites).
Hypoproteinemia (and hypoalbuminemia) can result from renal disease (albuminura) and
from bowel disease (gastroenteropathy).
Classes of plasma proteins under the different peaks:
Proteins of hemostasis (clotting factors, inhibitors, fibrinogen)
Immunoglobulins (IgG, IgA, IgD, IgE, IgM)
Complement proteins
Protease inhibitors (1-Antitrypsin, 2-Macroglobulin)
Binding/Transport proteins (for hormones, metal ions, hydrophobic molecules)
http://www.labtestsonline.org/understanding/analytes/electrophoresis/protein_groups.html
Major plasma proteins and their functions listed according to their electrophoretic group
GROUP
PROTEIN
FUNCTION
Albumin
Albumin
Main plasma protein; reduces tissue water

accumulation, transports many substances
Alpha1-globulin
Alpha1-Antitrypsin
Inactivates trypsin and other proteolytic

enzymes, reduces damage from inflammation
Orosomucoid
Immune response modifier, binds acidic drugs

such as lidocaine, steroid carrier
High Density Lipoprotein(HDL) Reverse transport of cholesterol

Alpha2-globulin
Beta-globulin
Alpha2-Macroglobulin Binds to and inactivates enzymes, preventing tissue

damage
Haptoglobin
Hemoglobin-binding protein, inhibits microbe iron uptake
Ceruloplasmin
Copper-containing enzyme, involved in normal iron

metabolism, inhibits microbe iron uptake
Transferrin
Iron transport and delivery to cells
Low Density Lipoprotein (LDL) Cholesterol delivery to tissue

Complement component 3 Helps regulate inflammatory response to foreign
substances
Gamma-globulin
IgA
Immunoglobulin involved in secretions
IgG
IgM
C-reactive protein
Fibrinogen
Major immunoglobulin; long term immunity

Initial response immunoglobulin
Inflammatory response mediator
Coagulation factor (found only in plasma, not serum)
Plasma electrophoretogram in some pathological cases:
Figure 3.21, Devlin. Examples of the Electrophoretic Mobility Patterns Observed For a
Normal Individual and Patients With Abnormal Concentrations of Serum Proteins,
Analyzed by Agarose Gel Electrophoresis.
a) Normal pattern
b) Immediate response (acute phase response): Stress or inflammation caused by
infection, injury or surgical trauma causes selective increase in haptoglobins in
alpha2 mobility band.
Acute-phase proteins that increase are: haptoglobin, alpha-2 macroglobulin,
ceruloplasmin, C-reactive protein, complement factors, certain blood clotting
factors
Acute-phase proteins that decrease are: albumin, antithrombin, transcortin
c) A late response pattern (chronic inflammation) associated with infection shows an
increase in the gamma globulin peak due to increase in immunoglobulins. Alpha2
band is also increased and albumin peak is decreased (as in acute phase response).
d) Immunosuppressive diseases cause a decrease in the gamma globulin peak.
e) Hepatic cirrhosis elevates gamma globulin with reduction in albumin. Albumin is made
by the liver, when decreased, gamma globulins are elevated to keep osmotic
pressure of the blood close to normal.
f) Monoclonal gammopathy, due to clonal synthesis of a unique immunoglobulin, sharply
increases the gamma globulin band. In multiple myeloma, neoplastic cells circulate in
blood, characterized by uncontrolled proliferation of the cells and secretion of
homogeneous gamma globulin, called M-protein.
g) Nephrotic syndrome shows a reduction in low molecular weight proteins that leave the
blood through the kidney. High alpha 2 band is present because of inflammation and
stress.
h) Protein-losing enteropathy shows loss of albumin and gamma globulins; slight
increase in the alpha2 band is due to a stressful stimulus seen in b) and g).
10

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Blood, Plasma and Serum Components

Gabriella Cs-Szabo, Ph.D.

BLOOD, PLASMA AND SERUM COMPONENTS

Human circulatory system

Gas exchange in alveoli

The circulatory system

3. What are the major components of blood?

A. Major functions of the cells:

Electron Microscopy Facility at The National

Courtesy: Department of Histology, Jagiellonian

3. What is plasma and serum?

Human plasma constituents by volume.

Scheme of a blood sample

Blood samples, right: freshly drawn;

4. Why do we use blood tests?

Figure 3.20. Devlin Electrophoresis pattern for plasma proteins at pH 8.6.

Examples of plasma proteins:

Main plasma protein; reduces tissue water

Inactivates trypsin and other proteolytic

Immune response modifier, binds acidic drugs

High Density Lipoprotein(HDL) Reverse transport of cholesterol

Alpha2-Macroglobulin Binds to and inactivates enzymes, preventing tissue

Hemoglobin-binding protein, inhibits microbe iron uptake

Copper-containing enzyme, involved in normal iron

Iron transport and delivery to cells

Low Density Lipoprotein (LDL) Cholesterol delivery to tissue

Immunoglobulin involved in secretions

Major immunoglobulin; long term immunity

Plasma electrophoretogram in some pathological cases:

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