Mar 1, 1987

Comparison of horseradish peroxidase and alkaline phosphatase-labelled antibodies in enzyme immunoassays

Annals of Clinical Biochemistry
K BeyzaviR Clift

Abstract

The periodate method was found to be most effective for preparing horseradish peroxidase-sheep anti-human and horseradish peroxidase-donkey anti-mouse immunoglobulin (IgG) conjugates. The conjugates were improved by carrying out the oxidation of the enzyme at low pH. Anti-human and anti-mouse IgG-peroxidase conjugates (0.5 mg/mL IgG and 0.7 mg/mL IgG, respectively) were used at 1:15,000 and 1:8000 dilutions, respectively, in a sandwich ELISA to detect human and mouse IgG in buffer or in a growth medium containing 20% foetal calf serum. Using the peroxidase conjugates, it was possible to detect human and mouse IgG at concentrations as low as 1 ng/mL. The glutaraldehyde method was found to be much more effective than the periodate method for conjugating alkaline phosphatase to the antibodies. The optimum dilutions for anti/human and anti-mouse IgG-alkaline phosphatase conjugates (0.18 mg/mL IgG and 0.3 mg/mL IgG, respectively) in ELISA were 1:500 and 1:1000, respectively. The detection limit with alkaline phosphatase conjugates was 7 ng/ml for human IgG and 4 ng/ml for mouse IgG.

  • References12
  • Citations8

Mentioned in this Paper

Ovoperoxidase
Alpha-Peroxidase
Structure of Calf of Leg
Immunoglobulin Activity
Cattle calf (organism)
Peroxidases
Equus asinus
Alkaline Phosphatase Measurement
Bos taurus
Alkaline Phosphatase

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