A novel monoclonal antibody-based enzyme-linked immunosorbent assay to determine luteinizing hormone in bovine plasma.
Domest Anim Endocrinol
; 48: 145-57, 2014 Jul.
Article
in En
| MEDLINE
| ID: mdl-24906940
The development of a novel enzyme-linked immunosorbent assay (ELISA) for determining luteinizing hormone (LH) in bovine plasma is described. Anti-bovine LH (bLH) monoclonal antibodies (mAbs) were produced and characterized. One mAb recognizing the bLH ß subunit was used for immunoaffinity purification of substantial amounts of biologically active bLH from pituitary glands. The purified bLH in combination with 2 anti-bLH ß subunit mAbs was used to develop a sandwich ELISA, which satisfied all the criteria required to investigate LH secretory patterns in the bovine species. The ELISA standard curve was linear over the range 0.05 to 2.5 ng/mL, and the assay proved suitable for measuring bLH in plasma without any prior treatment of samples. Cross-reactivity and recovery tests confirmed the specificity of the method. The intra- and inter-assay coefficients of variation ranged between 3.41% and 9.40%, and 9.29% and 15.84%, respectively. The analytical specificity of the method was validated in vivo by provocative tests for LH in heifers, using the LH releasing peptide gonadotropin-releasing hormone. In conclusion, the adoption of mAbs for this ELISA for coating the wells and labeling, combined with the easy one-step production of reference bLH, ensures long-term continuity in large-scale measurements of LH in the bovine species.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Cattle
/
Enzyme-Linked Immunosorbent Assay
/
Luteinizing Hormone
/
Antibodies, Monoclonal
Type of study:
Diagnostic_studies
Limits:
Animals
Language:
En
Journal:
Domest Anim Endocrinol
Journal subject:
ENDOCRINOLOGIA
/
MEDICINA VETERINARIA
Year:
2014
Document type:
Article
Country of publication:
United States