Development of a specific fluoroimmunoassay for ovine albumin.
A fluoroimmunoassay has been developed to measure serum levels of albumin in sheep. It employs ovine albumin labelled with fluorescein as the tracer and a rabbit antiserum raised against ovine albumin. Separation of the antibody bound and free fractions is achieved using a second antiserum directed against the Fc of rabbit immunoglobulin G and, to simplify the assay, the two antisera are premixed prior to use. Assay validation parameters are satisfactory and the reagents are predicted to be stable for at least one year at 4 degrees C. In contrast to the bromocresol green method, the assay is unaffected by immunoglobulins. A reference range for serum albumin levels has been established in lambs, normal ewes and ewes undergoing immunisation. Mean serum levels were 38.8, 51.3 and 37.8 g/l respectively. The sensitivity of the assay also enabled its use to monitor albumin levels at various stages during the production of specific antibody fragments from ovine antisera for therapeutic purposes.
Hreczuk-Hirst
D H
DH
Therapeutic Antibodies Ltd., Medical College of St Bartholomew's Hospital, London.
Harrison
K
K
Sidki
A
A
D000818:Animals; D001961:Bromcresol Green; D004355:Drug Stability; D015200:Fluoroimmunoassay / Q000379:methods* / Q000592:standards / Q000662:veterinary; D007074:Immunoglobulin G / Q000032:analysis; D007202:Indicators and Reagents; D012015:Reference Standards; D012016:Reference Values; D015203:Reproducibility of Results; D012709:Serum Albumin / Q000032:analysis* / Q000276:immunology / Q000592:standards; D012756:Sheep; D013053:Spectrophotometry
D007074:Immunoglobulin G; D007202:Indicators and Reagents; D012709:Serum Albumin; D001961:Bromcresol Green