Development of a Highly Sensitive Enzyme Immunoassay for Human Calcitonin Using Solid Phase Coupled with Multiple Antibodies
| Autor: | Yoshihiro Ashihara, Mitsuo Isomura, Atsuko Kawada, Nobuo Honda, Hiromasa Suzuki |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Calcitonin
030213 general clinical medicine medicine.medical_specialty medicine.drug_class Clinical Biochemistry 030209 endocrinology & metabolism Peptide Cross Reactions Monoclonal antibody Sensitivity and Specificity law.invention Immunoenzyme Techniques Mice 03 medical and health sciences 0302 clinical medicine law Internal medicine medicine Animals Humans Avidity Chemiluminescence chemistry.chemical_classification biology medicine.diagnostic_test Antibodies Monoclonal Reproducibility of Results General Medicine Molecular biology Rats Endocrinology chemistry Immunoassay biology.protein Antibody Quantitative analysis (chemistry) hormones hormone substitutes and hormone antagonists |
| Zdroj: | Annals of Clinical Biochemistry: International Journal of Laboratory Medicine. 36:629-635 |
| ISSN: | 1758-1001 0004-5632 |
| DOI: | 10.1177/000456329903600511 |
| Popis: | We have developed a highly sensitive chemiluminescent enzyme immunoassay for human calcitonin using three different mouse monoclonal antibodies that recognize the N-terminal, C-terminal and central portions of a human calcitonin molecule. The assay signal in a two-step sandwich enzyme immunoassay for human calcitonin using a solid phase coupled with a mixture of monoclonal antibodies, CT08 and OCT1, was 3·7-fold higher than when using either or both solid phases coupled with CT08 or OCT1, respectively. This enhancement is the result of improved avidity of immobilized antibodies and greater stability of the complex of immobilized antibodies and calcitonin in the first reaction, which resulted in greater reactivity of the immunocomplex with alkaline phosphatase-conjugate in the second reaction. The present assay showed a linear response up to 2·5 μg/L of human calcitonin and a high specificity for human calcitonin, but not for rat calcitonin, human calcitonin gene-related peptide and rat calcitonin gene-related peptide. The detection limit of human calcitonin was estimated to be 0·29 ng/L at zero (assay blank) + 3 SD. Interbatch coefficients of variation ranged from 2·2–26·7%. |
| Databáze: | OpenAIRE |
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