Development of a dot-blot assay for screening monoclonal antibodies to low-molecular-mass drugs
| Autor: | Yon-Yih Chen, Rad K. Nair, Sushil D. Rege, Larissa Coombs Harwick, Gangamani S. Beligere, Jonathan Grote, Kevin R. Rupprecht, Jeffrey R. Fishpaugh, Zhen Lin |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Hydrocortisone
medicine.drug_class medicine.medical_treatment Immunoblotting Antibiotics Anti-Inflammatory Agents Biophysics Dot blot Monoclonal antibody Biochemistry Tacrolimus Steroid chemistry.chemical_compound medicine Molecular Biology Sirolimus Primary (chemistry) Chromatography Molecular mass Antibodies Monoclonal Collodion Reproducibility of Results Cell Biology Anti-Bacterial Agents Carbamazepine Methotrexate Pharmaceutical Preparations chemistry Cyclosporine Acridines Anticonvulsants Gentamicin Gentamicins Nitrocellulose Immunosuppressive Agents medicine.drug |
| Zdroj: | Analytical Biochemistry. 407:160-164 |
| ISSN: | 0003-2697 |
| Popis: | Dot-blot is a versatile and simple analysis to perform. We adapted this method as a simple identity test for monoclonal antibodies to a number of small compounds: three transplant drugs, an anticonvulsant, a steroid, an anticancer drug, and an antibiotic. Immunology-based identity tests using low-molecular-mass organic compounds have historically been a challenge to develop. We modified the traditional dot-blot assay to serve as an identity test for monoclonal antibodies to carbamazepine, sirolimus, tacrolimus, cyclosporine, cortisol, methotrexate, and gentamicin. The primary obstacle was the immobilization of these organic compounds on nitrocellulose as nitrocellulose is also soluble in most of the organic solvents in which the compounds are soluble. We evaluated different membranes, solvents, and chemical forms of these organic compounds to overcome this challenge. A number of incubation and washing solutions were also investigated. By varying the chemical form, concentration, and incubation conditions, a set of effective and reproducible identity tests were developed for these monoclonal antibodies. |
| Databáze: | OpenAIRE |
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