1.

Isoenzyme name Composition Electrophoretic Present in Elevated in

migration
LDH 1 HHHH (H4) Fastest moving Myocardium, RBC, Myocardial
Kidney Infarction
LDH 2 HHHM (H3M1) Myocardium, RBC, Kidney disease,
Kidney Megaloblastic
Anemia
LDH 3 HHMM (H2M2) Brain Leukemia,
Malignancy
LDH 4 HMMM (H1M3) Lung, Spleen Pulmonary
Infarction
LDH 5 MMMM (M4) Slowest Moving Skeletal Muscle, Skeletal muscle
Liver and Liver diseases

2.

A. Electrophoresis

Due to their different amino acid compositions the LDH isoenzymes can be separated by electrophoretic
methods (agarose, cellulose-acetate, polyacrylamide can be used as a matrix). In an electric field at pH
8.8 the LDH1 migrates at the fastest rate towards the anode, while the LDH5 is the slowest isoenzyme.
After electrophoresis the LDH isoenzymes can be visualized by an activity staining process where the
product of the enzymatic reaction is a water-insoluble dye precipitating in the gel where the LDH
enzymes are located.

B. Substrate Specificity

The LDH isoenzymes also show different substrate specificities towards pyruvate and 2-oxobutyrate as
substrates. While LDH1 shows a higher specific activity when 2-oxobutyrate is used, the LDH5 isoenzyme
is more active with pyruvate. The ratio of LDH activities measured in serum samples using the two
substrates (HBDH/LDH) varies between 0.63-0.81 in normal cases. Ratios higher than 0.83 are
characteristic for a myocardial infarction. Liver diseases are diagnosed when the ratio of enzyme
activities is lower than 0.61.

3.

LDH may be used as a general indicator for the severity of acute and chronic tissue damage.
LDH may be used to detect and monitor progressive conditions such as anemia including
hemolytic anemia, megaloblastic anemia and severe infections.
LDH determine prognosis or monitor treatment i.e. chemotherapy of cancer such as germ cell
tumors e.g. testicular and ovarian cancer, lymphoma, leukemia and neuroblastoma.
4.

Pyruvate Kinetic Method

5.

*Hemolysis- affects both total LD and LD1/LD2 ratio

*Contact with the clot increases LD

*Physical agitation of specimens, as occurs in most pneumatic tube systems, tends to cause some
hemolysis and increased LD

*Granulocyte-macrophage colony stimulating factor appears to increase LD in parallel to the increase in

WBC count

* Exercise causes, at most, slight increases in total LD