Development and validation of a biosensor-based immunoassay for progesterone in bovine milk

• Published in: Journal of immunological methods 2002-09, Vol.267 (2), p.131-138
• Main Authors: Gillis, Els H, Gosling, James P, Sreenan, Joseph M, Kane, Marian
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal; Biological and medical sciences; Biosensor assays; Biosensors; Biotechnology; Cattle; Dairying; Enzyme-Linked Immunosorbent Assay; Estrus Detection; Female; Fundamental and applied biological sciences. Psychology; Heat detection; Immunoassay - methods; Immunoassay - statistics & numerical data; Methods. Procedures. Technologies; Milk - chemistry; Mother. Fetoplacental unit. Mammary gland. Milk; Pregnancy; Pregnancy. Parturition. Lactation; Progesterone; Progesterone - analysis; Progesterone - immunology; Reproducibility of Results; Reproduction; SPR; Surface Plasmon Resonance - methods; Surface Plasmon Resonance - statistics & numerical data; Various methods and equipments; Vertebrates: reproduction
• ISSN: 0022-1759; 1872-7905
• DOI: 10.1016/S0022-1759(02)00166-7

We have developed a rapid automated immunoassay, using the BIACORE™ surface plasmon resonance (SPR) biosensor, to measure progesterone in bovine milk. The assay was designed as an inhibition assay with progesterone covalently immobilised to the carboxymethyl dextran matrix of a CM5 sensor chip. A fixed amount of monoclonal anti-progesterone antibody 39C5H7 was mixed 9:1 with the sample and the amount of free antibody was then determined using biomolecular interaction analysis (BIA) by injection of the mixture over the immobilised progesterone sensor surface. The assay was designed to cover the concentration range 0.5 to 50 ng/ml. The limit of detection (LOD) was 3.56 ng/ml. Reproducibility of the assay was very good with both intra-assay and inter-assay coefficients of variation