Serum Proteins

1- Total protein
2- Albumin
3- Globulin

Introduction:
• Amino acids are major metabolic intermediates and the basic
structural units of proteins.
• Amino acids are organic compounds containing both an
amino group (—NH2) and a carboxyl group (—COOH).
• Amino acids that are present in proteins are α-amino acids,
with amino groups linked to the α-carbon, as in diagram:

• R represents a variety of different sidechains.

Plasma Proteins:
• Human plasma contains a complex mixture of proteins.

• About 100 of the most abundant plasma proteins have been

recognized, but the physiological functions of a small
proportion of them have been known.

• The most abundant proteins are those synthesized by the

liver and secreted directly into the circulation, and
immunoglobulins which are synthesized and secreted by B-
lymphocytes (plasma cells).

• Protein in the serum is made up of albumin and globulin. The

globulin in turn is made up of α1, α2, β, and γ globulins.

• Whereas in the plasma; in addition to the albumin and

globulins; fibrinogen and the other clotting factors (which are
proteins) are also present.

Clinical significance:
Relative hypoproteinemia:
Occurs when there is an increase in plasma water volume
(hemo-dilution) such as in:
1.Water intoxication (psychological).
2.Massive infusion of intravenous fluid
Absolute hypoproteinemia:
Absolute hypoproteinemia is due to low level of albumin in the
blood. It is common and may be caused by:
1.Low dietary intake of protein e.g. malnutrition, starvation.
2.Decreased synthesis of protein e.g. severe liver disease
(cirrhosis).
3.Increased loss of protein from the body:
a- Renal loss of protein e.g. nephrotic syndrome.
b- Gastrointestinal loss of protein e.g. protein losing
enteropathy.
Relative hyperproteinemia:
Occurs when there is a decrease in the volume of plasma water
(hemo-concentration) such as in dehydration which may be
caused by:
1. Severe vomiting
2. Severe diarrhea
3. Diabetic ketoacidosis.

Absolute hyperproteinemia:
Absolute hyperproteinemia may be mild or marked.

Mild hyperproteinemia may be caused by an increase in the

concentration of specific proteins e.g. in infection (there is
increased production of gamma globulins).

Marked hyperproteinemia may be caused by high levels of

monoclonal immunoglobulins produced in multiple myeloma
and other malignant paraproteinemia's.

Laboratory measurement of plasma proteins:

1- Total Protein
Specimen collection:
- Serum or plasma can be used.

- Avoid prolonged application of tourniquet, which causes

hemoconcentration leading to spurious increase in
concentration of all plasma proteins.

Principle of the test:

The most widely used method of measuring serum protein is
the Biuret reaction. It is colorimetric method in which
there is reaction between the peptide bonds of protein and
cupric ions (Cu+2) in alkaline solution to form a colored
complex. The absorbance of the complex is directly
proportional to the concentration of total protein in
specimen. The absorbance is measured at 550 nm.
Reagents:

- Biuret reagent:
It contains:

1. Sodium potassium tartarated (to complex cupric ions and

maintain their solubility in alkaline solution).
2. Sodium hydroxide
3. Potassium iodide
4. Copper sulfate

- Standard: 6 g/dl.

Manual Procedure:
Pipette into well identified
Blank Standard Test
tubes
Biuret reagent 1 ml 1 ml 1 ml
Distilled water 20 µl
Standard 20 µl
Specimen 20 µl
- Mix well.
- Put the tubes in the rack and let stand for 10 minutes at
room temperature.
- Record absorbance at 550 nm.
Calculation:
𝐀𝐛𝐬 𝐓𝐞𝐬𝐭 − 𝐀𝐛𝐬 𝐁𝐥𝐚𝐧𝐤
𝐂𝐨𝐧𝐜. 𝐨𝐟 𝐓𝐨𝐭𝐚𝐥 𝐩𝐫𝐨𝐭𝐞𝐢𝐧 = 𝐗 𝐒𝐭𝐝 𝐂𝐨𝐧𝐜.
𝐀𝐛𝐬 𝐒𝐭𝐝 − 𝐀𝐛𝐬 𝐁𝐥𝐚𝐧𝐤

A = absorbance, measured by spectrophotometer.

Standard concentration = 6 g/dl

Expected values:

for adults: 6.0 - 8.0 g/dl