| Autor: |
Thurmond LM; Division of Bioanalysis and Drug Metabolism, Glaxo Wellcome Inc., Research Triangle Park, NC 27709, USA., Reese MJ, Donaldson RJ, Orban BS |
| Jazyk: |
angličtina |
| Zdroj: |
Journal of pharmaceutical and biomedical analysis [J Pharm Biomed Anal] 1998 Apr; Vol. 16 (8), pp. 1317-28. |
| DOI: |
10.1016/s0731-7085(97)00168-4 |
| Abstrakt: |
A kinetic enzyme immunoassay was developed and validated to quantitate human antibodies to the humanized monoclonal antibody CAMPATH1-1H (C1H) in human serum. The assay was configured using C1H-coated 96-well plates which were blocked with bovine serum albumin, and incubated with dilutions of human serum containing anti-C1H antibody. Antibody was detected using biotinylated C1H followed by streptavidin-conjugated alkaline phosphatase and p-nitrophenyl phosphate. Absorbance data were collected for 10 min, and mOD min-1 data were exported to MultiCalc data analysis software. A 4-parameter logistic-log algorithm was shown to model the data through the range of the standard curve within 15% of nominal values. The overall assay performance coefficient of variation by ANOVA was 9.2%. The lower limit of detection was defined at 160 Units ml-1. The anti-idiotype antibody standard stock solution is stable at 4 degrees C and at -80 degrees C for at least 11 months in buffer. The anti-idiotype antibody controls are stable for at least seven freeze-thaw cycles and at least 6 months in human serum stored at -20 degrees C. A strategy was devised by which to establish the specific antibody potency for any given batch of anti-C1H antibody standard relative to the Reference Standard. This EIA has been used to quantify and characterize anti-C1H antibody in human serum in support of clinical safety and efficacy studies. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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