An indispensable part of the production process of recombinant human α2a/α2b interferon is the quality control of the product, and the bioactivity of interferon is usually detected by the HEp-2/VSV or Wish/VSV system. This system is often not stable enough due to the influence of VSV or cells, and the assay period is long. (Source: Laboratory Guidelines for Basic Medical Immunology, edited by Jin Boquan and Li Enshan, 1st edition, Beijing: World Book Publishing House, 1990).

Operation method

Sandwich ELISA

Principle

Two McAb against different epitopes of rHuIFN-α2a and rHuIFN-α2b molecules were selected, one McAb as the coating antibody and the other McAb labeled horseradish peroxidase (HRP) as the conjugate, which catalyzes the color development of the substrate. The standard curve was plotted according to the OD value of the standard, and the content of the specimen to be examined was found on the standard curve.

Materials and Instruments

Coated Antibodies Enzyme-Labeled Antibodies Standards ABTS Substrates
Buffer Wash Solution Diluent Terminator
ELISA plates

Move

1. Add 100 μl of diluted antibody into the ELISA plate. Leave at 4 ℃ for 24～48 h. 2.
2. Wash the plate 3 times and add 10-fold serial dilutions of the specimen to be tested (4 dilutions are sufficient) or doubly diluted standards (6 dilutions). Wash 3 times, add 10-fold serial dilution of the specimen to be tested (4 dilutions can be done) or doubly diluted standard (6 dilutions) 100 μl/well. 37 ℃, 1 h
3. Wash 3 times, add 100 μl/well of diluted enzyme antibody. 37 ℃, 45 min
4. Wash 4 times, add 100 μl/well of prepared ABTS substrate solution. RT, 30 min
5. Measure 410 nm OD 6. Plot standard curve

Caveat

1. The specimen to be examined must be free of contamination and hemolysis.

2. When the specimen is diluted and added to the ELISA plate, take care to add the specimen sequentially from low concentration to high concentration.

3. prohibit the addition of sodium azide to the conjugate diluent and substrate buffer.

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