title: A simple monoclonal antibody based ELISA for free protein S. Comparison with PEG precipitation.
abstract: A simple monoclonal antibody based ELISA for free Protein S, compatible with out existing ELISA for total Protein S has been developed, and its performance compared with the conventional PEG precipitation method of free Protein S assay. The normal range (mean +/- 2 SD) was 0.19-0.54 iu/ml free Protein S. The mean intra assay variation was 5.24% and the mean inter assay variation was 5.50%. A total of 102 routine diagnostic samples from patients referred for prothrombotic investigation (six assays for each method) were assayed by PEG precipitation (mean 0.32 iu/ml, SD 10.60), and the monoclonal ELISA (mean 0.34, SD 0.9). Paired t-test analysis of the two data sets indicated no significant difference between them (P < 0.001). In this sample population, there was no significant difference in free Protein S values when assayed by monoclonal based ELISA or by PEG precipitation. The monoclonal assay has proved to be reliable, accurate and precise. The monoclonal ELISA is simpler, quicker and easier to perform in routine use. Data generated is directly comparable to that generated by PEG precipitation. This methodology would be suitable for laboratories currently measuring Protein S by ELISA.
authors.affiliation: Diagnostic Laboratory, Roy Free Hampstead NHS Trust, Royal Free Hospital, London, UK.
authors.forename: P J
authors.initials: PJ
authors.lastname: Murdock
authors.forename: S
authors.initials: S
authors.lastname: Brooks
authors.forename: G
authors.initials: G
authors.lastname: Mellars
authors.forename: G
authors.initials: G
authors.lastname: Cheung
authors.forename: D
authors.initials: D
authors.lastname: Jacob
authors.forename: D L
authors.initials: DL
authors.lastname: Owens
authors.forename: M
authors.initials: M
authors.lastname: Parmar
authors.forename: A
authors.initials: A
authors.lastname: Riddell
mesh_terms: D000704:Analysis of Variance; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D011092:Polyethylene Glycols; D011233:Precipitin Tests; D017293:Protein S / Q000032:analysis* / Q000276:immunology* / Q000378:metabolism; D015203:Reproducibility of Results
chemical_list: D000911:Antibodies, Monoclonal; D017293:Protein S; D011092:Polyethylene Glycols