Competitive ELISA of Bovine Lactoferrin with Bispecific Monoclonal Antibodies | Zendy

Hiroshi Shinmoto | Zendy; Masuko Kobori | Zendy; Tojiro Tsushida | Zendy; Kazuki Shinohara | Zendy

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Competitive ELISA of Bovine Lactoferrin with Bispecific Monoclonal Antibodies

Author(s) -

Hiroshi Shinmoto,

Masuko Kobori,

Tojiro Tsushida,

Kazuki Shinohara

Publication year - 1997

Publication title -

bioscience biotechnology and biochemistry

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.509

H-Index - 116

eISSN - 1347-6947

pISSN - 0916-8451

DOI - 10.1271/bbb.61.1044

Subject(s) - monoclonal antibody , horseradish peroxidase , affinity chromatography , chemistry , chromatography , lactoferrin , antibody , antigen , immunoassay , microbiology and biotechnology , enzyme , biochemistry , biology , immunology

A mouse hybrid-hybridoma, HH1-4-3, secreting IgG1 class bispecific antibodies to bovine lactoferrin (bLF) and horseradish peroxidase (HRPO), was established previously. The competitive enzyme-linked immunosorbent assay (ELISA) of bLF using the HH1-4-3 culture supernatant was not sensitive enough to measure bLF concentration in biological fluids. To improve the sensitivity of the competitive ELISA, we fractionated the bispecific antibodies by antigen affinity column chromatography. A column immobilized with bLF adsorbed 60% of the antibodies of the HH1-4-3 supernatant, and the amount of antibodies adsorbed on a column immobilized with HRPO was less than 5%. The competitive ELISA of bLF using the affinity purified bispecific antibodies through an HRPO-immobilized column chromatography showed a good standard curve at bLF concentrations of 10 ng/ml to 100 micrograms/ml.

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